Methods and compositions for treating intestinal disorder

ABSTRACT

The presently disclosed subject matter relates to method, compositions and food products for improving intestinal health, treating intestinal dysbiosis and/or treating an intestinal disorder in a subject, e.g., a human or a companion animal.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to U.S. Provisional Application No. 62/796,021, filed on Jan. 23, 2019, which is incorporated herein by reference in its entirety.

FIELD

The presently disclosed subject matter relates to method, compositions and food products for improving intestinal health, treating intestinal dysbiosis and/or treating an intestinal disorder in a subject, e.g., a human or a companion animal.

BACKGROUND

Inflammatory bowel disease (IBD), including Crohn's disease and ulcerative colitis, is a multi-factorial and debilitating disease characterized by chronic immune-pathology, disruption of intestinal homeostasis and altered composition of the gut microbiome (dysbiosis). Several lines of evidence point to resident gut bacteria as important factors in the etiology of IBD. First, disease is often more severe in areas of the intestine with the highest microbial biomass, and antibiotics are frequently used as an adjunct therapy with immunosuppressants or monoclonal antibodies for managing IBD^(1,2). Second, genome-wide associations studies have identified numerous susceptibility loci in genes responsible for recognizing or responding to bacteria³. Finally, in some mouse models of colitis, disease can be transferred to naive hosts via fecal transplant⁴⁻⁶, suggesting a causal role for gut microbes in disease. Collectively, these findings have led to a ‘two-hit’ model for IBD in which both host genetics and microbial factors influence disease presentation, highlighting an opportunity to develop novel treatments for IBD that target the microbiome. Thus, there is a need for novel methods and compositions for treating IBD and other intestinal disorders that target gut microbiome and metabolites thereof.

SUMMARY OF THE INVENTION

The presently disclosed subject matter provides a pharmaceutical composition, dietary supplement and functional food for medicament. In certain embodiments, the pharmaceutical composition, dietary supplement or functional food comprises an effective amount of a bacterium capable of producing a first bile acid for use as a medicament. In certain embodiments, the pharmaceutical composition, dietary supplement or functional food further comprises an effective amount of a second bile acid. In certain embodiments, the pharmaceutical composition, dietary supplement or functional food is for the treatment of an intestinal disorder in a subject in need thereof.

In certain embodiments, the bacterium comprises a bile acid-inducible operon (bai operon). In certain embodiments, the bile acid-inducible operon (bai operon) comprises a nucleotide sequence that is at least about 90% homologous or identical to SEQ ID NO: 1 or 3, or any functional fragment thereof. In certain embodiments, the bile acid-inducible operon (bai operon) comprises the nucleotide sequence set forth in SEQ ID NO: 1 or 3.

In certain embodiments, the bacterium comprises a 16s rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to SEQ ID NO: 2 or 4. In certain embodiments, the bacterium comprises a 16s rRNA comprising the nucleotide sequence set forth in SEQ ID NO: 2 or 4.

In certain embodiments, the bacterium is C. hiranonis, C. scindens or combination thereof. In certain embodiments, the bacterium is C. hiranonis.

In certain embodiments, the first bile acid and/or the second bile acid is selected from the group consisting of chenodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, taurochenodeoxycholic acid, taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid and any combination thereof. In certain embodiments, the first bile acid and/or the second bile acid is a secondary bile acid. In certain embodiments, the secondary bile acid is selected from the group consisting of taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid and any combination thereof. In certain embodiments, the secondary bile acid is deoxycholic acid and/or lithocholic acid.

In certain embodiments, the subject is a dog. In certain embodiments, the intestinal disorder is an acute enteropathy or a chronic enteropathy. In certain embodiments, the chronic enteropathy is selected from the group consisting of food responsive enteropathy, antibiotic responsive enteropathy, and idiopathic inflammatory bowel disease (IBD). In certain embodiments, the intestinal disorder is idiopathic inflammatory bowel disease (IBD).

In certain embodiments, the bacterium is transformed with a vector comprising a bile acid-inducible operon (bai operon). In certain embodiments, the bacterium is selected from the genus of Clostridium.

In certain embodiments, the amount of the bacterium is between about 10 thousand CFU and about 100 trillion CFU. In certain embodiments, the second bile acid is between about 10 mg/unit dose and about 500 mg/unit dose.

In certain embodiments, the first bile acid and the second bile acid are the same. In certain embodiments, the first bile acid and the second bile acid are different.

The presently disclosed subject matter provides C. hiranonis for use as a functional food or supplement to prevent onset of a GI condition or as a medicament. In certain embodiments, the C. hiranonis is for the treatment of an intestinal disorder in a subject in need thereof. The presently disclosed subject matter provides C. scindens functional food or supplement to prevent onset of a GI condition or for use as a medicament. In certain embodiments, the C. scindens is for the treatment of an intestinal disorder in a subject in need thereof.

The presently disclosed subject matter provides deoxycholic acid for the treatment of inflammatory bowel disease (IBD) in a subject in need thereof. The presently disclosed subject matter provides lithocholic acid for the treatment of inflammatory bowel disease (IBD) in a subject in need thereof.

The presently disclosed subject matter provides a dietary supplement or a food product comprising an effective amount of a bacterium capable of producing a first bile acid. In certain embodiments, the dietary supplement or a food product further comprises an effective amount of a second bile acid. In certain embodiments, the food product improves intestinal health in a subject. In certain embodiments, the amount of the bacterium is between about 10 thousand CFU and about 100 trillion CFU. In certain embodiments, the second bile acid is between about 100 mg/daily serving dose and about 1000 mg/daily serving dose.

In certain embodiments, the food product is a pet food product. In certain embodiments, the food product is a dog food product.

The presently disclosed subject matter provides a method of treating an intestinal disorder in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition, dietary supplement or functional food disclosed herein, an effective amount of a food product disclosed herein, or combination thereof. In certain embodiments, the method further comprises monitoring an intestinal microorganism in the subject. In certain embodiments, the intestinal microorganism is sampled from a fecal sample of the subject.

The presently disclosed subject matter provides a method for determining susceptibility of an intestinal disorder in a companion animal. In certain embodiments, the method comprises:

a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal;

b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and

c) determining that the companion animal is susceptible of an intestinal disorder, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HQ802983.1.1440, GQ449092.1.1375, GQ448744.1.1393, KF842598.1.1394, HG798451.1.1400, New.ReferenceOTU52, HK555938.1.1357, FJ957494.1.1454, FN667392.1.1495, New.ReferenceOTU54, HQ760911.1.1437, GQ006324.1.1342, FJ950694.1.1472, FM865905.1.1392, FJ506371.1.1371, FJ957528.1.1445, JF712675.1.1540, New.ReferenceOTU82, AB009242.1.1451, HQ751549.1.1448, AB506370.1.1516, DQ057365.1.1393, FN667422.1.1495, AJ270486.1.1241, FN668375.4306350.4307737, GQ867426.1.1494, GX182404.8.1529, JF224013.1.1362, GQ448246.1.1389, KC245406.1.1465, FN667084.1.1493, EU470512.1.1400, EU768569.1.1352, AY239462.1.1500, KC504009.1.1465, FM179752.1.1686, New.ReferenceOTU114, HK557089.3.1395, JQ208181.1.1352, HQ803964.1.1435, AM276759.1.1484, JN387556.1.1324, GQ448486.1.1387, HK694029.9.1487, HQ754680.1.1441, FN563300.1.1447, FP929060.3837.5503, GQ448506.1.1374, Enterococcus durans, C. perfringens, or E. coli.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of HQ802983.1.1440, GQ449092.1.1375, GQ448744.1.1393, KF842598.1.1394, HG798451.1.1400, New.ReferenceOTU52, HK555938.1.1357, FJ957494.1.1454, FN667392.1.1495, New.ReferenceOTU54, HQ760911.1.1437, GQ006324.1.1342, FJ950694.1.1472, FM865905.1.1392, FJ506371.1.1371, FJ957528.1.1445, JF712675.1.1540, New.ReferenceOTU82, AB009242.1.1451, HQ751549.1.1448, AB506370.1.1516, DQ057365.1.1393, FN667422.1.1495, AJ270486.1.1241, FN668375.4306350.4307737, GQ867426.1.1494, GX182404.8.1529, JF224013.1.1362, GQ448246.1.1389, JF807116.1.1260, KC245406.1.1465, FN667084.1.1493, EU470512.1.1400, EU768569.1.1352, AY239462.1.1500, KC504009.1.1465, FM179752.1.1686, New.ReferenceOTU114, HK557089.3.1395, JQ208181.1.1352, HQ803964.1.1435, AM276759.1.1484, JN387556.1.1324, GQ448486.1.1387, HK694029.9.1487, HQ754680.1.1441, FN563300.1.1447, FP929060.3837.5503, GQ448506.1.1374, Enterococcus durans, C. perfringens, E. coli and any combination thereof. In certain embodiments, the first intestinal microorganism is C. perfringens, E. coli and any combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of EU774020.1.1361, HQ793763.1.1451, HQ792787.1.1438, New.ReferenceOTU109, HQ792778.1.1436, or DQ113765.1.1450.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of EU774020.1.1361, HQ793763.1.1451, HQ792787.1.1438, New.ReferenceOTU109, HQ792778.1.1436, DQ113765.1.1450, and any combination thereof.

In certain embodiments, the method further comprises providing a customized recommendation of a treatment regimen, and/or further monitoring the intestinal microorganism, when the first amount of the first intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.

The presently disclosed subject matter provides a method for determining responsiveness of a companion animal having an intestinal disorder to a diet. In certain embodiments, the method comprises:

a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal;

b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and

c) determining that the companion animal is responsive to the diet, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism, or determining that the companion animal is non-responsive to the diet, when the first amount of the intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, or JQ208053.1.1336.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of New.ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, JQ208053.1.1336, and any combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, or HK555938.1.1357.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, HK555938.1.1357, and any combination thereof.

In certain embodiments, the method further comprises administering the diet to the companion animal when companion animal is determined as responsive to the diet. In certain embodiments, the method further comprises administering the diet, a steroid and optionally an antibiotic to the companion animal when companion animal is determined as non-responsive to the diet.

In certain embodiments, the determination in step c) occurs before administering the diet or the diet, the steroid and optionally the antibiotic to the companion animal.

The presently disclosed subject matter provides a method for determining effectiveness of a diet for treating an intestinal disorder in a companion animal. In certain embodiments, the method comprises:

a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal after administering a diet to a companion animal for treating an intestinal disorder;

b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and

c) determining that the diet is effective for treating an intestinal disorder, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism, or determining that the diet is ineffective for treating an intestinal disorder, when the first amount of the intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK557089.3.1395, or GQ448336.1.1418.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of HK557089.3.1395, GQ448336.1.1418, and combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of KF842598.1.1394, GQ006324.1.1342, HQ802983.1.1440, JN387556.1.1324, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, or GQ448468.1.1366.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of KF842598.1.1394, GQ006324.1.1342, HQ802983.1.1440, JN387556.1.1324, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, GQ448468.1.1366, and any combination thereof.

In certain embodiments, the method further comprises administering the diet to the companion animal when companion animal is determined as responsive to the diet. In certain embodiments, the method further comprises administering the diet, a steroid and optionally an antibiotic to the companion animal when companion animal is determined as non-responsive to the diet.

In certain embodiments, the determination in step c) occurs before administering the diet or the diet, the steroid and optionally the antibiotic to the companion animal.

In certain embodiments, the reference amount of an intestinal microorganism derived from a mean amount of the intestinal microorganism in a plurality of healthy companion animals. In certain embodiments, the amount of the intestinal bacterium is measured from a fecal sample of the subject.

The presently disclosed subject matter provides a diet for increase a population of a bacterium capable of producing a bile acid in a companion animal. In certain embodiments, the diet comprises protein, fat, crude fiber, total dietary fiber, carbohydrate, calcium, phosphorus, sodium, chloride, potassium, magnesium, iron, copper, manganese, zinc, iodine, selenium, vitamin A, vitamin D3, vitamin E, vitamin C, thiamine (vitamin B1), riboflavin (vitamin B2), pantothenic acid, niacin, pyridoxine (vitamin B6), folic acid, biotin, cobalannin (vitamin B12), choline, arginine, lysine, methionine, cystine, taurine, linoleic acid, arachidonic acid, Omega-6 fatty acids, Omega-3 fatty acids, EPA, and/or DHA.

In certain embodiments, the subject is a dog. In certain embodiments, the diet is a Royal Canin Veterinary Diet. In certain embodiments, the diet is selected from the group consisting of Ultamino, Hydrolyzed Protein Adult HP Dry, Hydrolyzed Protein Wet, Hydrolyzed Protein Adult PS Dry, Hydrolyzed Protein Moderate Calorie Dry, Hydrolyzed Protein Small Dog Dry, Hydrolyzed protein Treats, and any combination thereof.

In certain embodiments, the bacterium comprises a bile acid-inducible operon (bai operon). In certain embodiments, the bacterium is C. hiranonis, C. scindens or combination thereof. In certain embodiments, the bacterium is C. hiranonis.

The presently disclosed subject matter provides a Royal Canin Veterinary Diet for the treatment of an intestinal disorder in a dog, wherein the dog comprises a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism, and wherein the first amount of the first intestinal microorganism is higher than a first reference amount of the first intestinal microorganism, and/or the second amount of the second intestinal microorganism is lower than a second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, or JQ208053.1.1336.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of New.ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, JQ208053.1.1336, and any combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, or HK555938.1.1357.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, HK555938.1.1357, and any combination thereof.

In certain embodiments, the Royal Canin Veterinary Diet is selected from the group consisting of Ultamino, Hydrolyzed Protein Adult HP Dry, Hydrolyzed Protein Wet, Hydrolyzed Protein Adult PS Dry, Hydrolyzed Protein Moderate Calorie Dry, Hydrolyzed Protein Small Dog Dry, Hydrolyzed protein Treats, and any combination thereof.

The presently disclosed subject matter provides a bile acid for the treatment of an intestinal disorder in a dog. In certain embodiments, the bile acid is selected from the group consisting of chenodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, taurochenodeoxycholic acid, taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid and any combination thereof. In certain embodiments, the bile acid is a secondary bile acid. In certain embodiments, the secondary bile acid is selected from the group consisting of taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid and any combination thereof. In certain embodiments, the secondary bile acid is deoxycholic acid and/or lithocholic acid.

The presently disclosed subject matter provides a kit comprising a presently disclosed pharmaceutical composition, dietary supplement, functional food, food product, diet or bile acid. In certain embodiments, the kit further comprises written instructions for treating and/or preventing an intestinal disorder.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1C depict that diet therapy induces rapid and durable remission in canine model of chronic enteritis. FIG. 1A is a schematic showing clinical study design for identifying diet responsive (DR) and non-diet responsive (NDR) dogs. Antibiotics (Abtx) and Prednisone (Pred) treatments are indicated. Abbreviated Canine Chronic Enteropathy Clinical Activity Index (CCECAI) scores were assessed at four different time points in DR (n=20) (FIG. 1B) and NDR (n=9) (FIG. 1C) animals. ns=not significant, ** p<0.01, p<0.0001 using Wilcoxon rank sum test.

FIGS. 2A-2F depict identification of microbial community profiles associated with treatment outcome. FIG. 2A is a ternary plot of phylum level OTUs from top 5 most abundant phyla among healthy (right), DR (left) and NDR (top) animals. Bubble size represents the log 2 OTU abundance. Relative abundance of E. coli (FIG. 2B) and C. perfringens (FIG. 2D) in animals with active disease (day 0) and healthy dogs. Spearman correlation between log 10 abundance of E. coli (FIG. 2C) or Clostridium sp. (FIG. 2E) and CCECAI disease score. FIG. 2F depicts differentially abundant OTUs between DR and DNR animals at day 0. Y-axis value represents the log 2 fold change for DR versus NDR. Arrow marks the OTU corresponding to C. perfringens. p<0.05, p<0.01 using Wilcoxon rank sum test (or Wilcoxon signed-rank test if available). Spearman correlations in panel C and E are significant (p<0.05) with correlation coefficients of 0.2109 and 0.2324, respectively.

FIGS. 3A-3F depict that therapeutic diet ameliorates dysbiosis associated with chronic enteritis. FIG. 3A depicts Pielou's evenness index for DR animals at different time points in the study. FIG. 3B depicts the principal coordinate analysis (PCoA) based on unweighted Unifrac distance for DR. FIG. 3C depicts the phylogenetic distance (unweighted Unifrac) to healthy controls for DR animals. FIG. 3D depicts the stream plot showing phylum level dynamics of microbiota structure for DR animals throughout the study. FIG. 3E depicts the volcano plot showing differentially abundant OTUs enriched in either DR dogs with active disease (day 0, red points) or in remission after diet therapy (day 14, blue points). Selected taxa (e.g., Escherichia-Shigella spp., Clostridium spp.) are labeled. The relative abundance of E. coli (FIG. 3F) and C. perfringens (FIG. 3G) in DR animals throughout the study and compared to healthy controls. ns=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon rank sum test (or Wilcoxon signed-rank test if paired data was available).

FIGS. 4A-4F depict diet-induced changes in the microbiome associated with remission. FIG. 4A depicts Pielou's evenness index in NDR animals, and their phylogenetic distance (unweighted Unifrac) to healthy dogs is shown in FIG. 4B. FIG. 4C depicts the stream plot showing phylum level dynamics of microbiota structure for NDR animals throughout the study. Diet therapy began at day 0, metronidazole administration at day 14, and prednisone at day 28 (see methods). FIG. 4D depicts the bubble plot showing differentially abundant genera (fold change >2 and P<0.05) between day 14 versus day 0 for DR (left) and NDR (right) animals. Bubble size indicates absolute log fold change between day 14 and day 0, and color reflects direction of change. FIGS. 4E and 4F depict the relative abundance of E. coli (FIG. 4E) and C. perfringens (FIG. 4F) in NDR animals throughout the study and compared to healthy controls. ns=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon rank sum test (or Wilcoxon signed-rank test if paired data was available).

FIGS. 5A-5H depict that diet-induced remission is associated with metabolic reprogramming and increased levels of secondary bile acids. FIG. 5A depicts a PCA analysis of KEGG pathways based on the results of Tax4Fun analysis. FIG. 5B depicts the first principal component (Dim 1) from panel A, for all time points. FIG. 5C depicts a heatmap showing the shift of metabolic potentials from fat/lipid metabolism to carbohydrate/sugar metabolism as DR animals receive diet therapy. FIG. 5D depicts the relative abundance of the KEGG pathway for secondary bile acid biosynthesis, predicted based on 16S sequence data. FIGS. 5E-5H depict the levels of deoxycholic (FIG. 5E) and lithocholic acid (FIG. 5F) measured in the stool of DR animals and NDR animals (FIGS. 5G and 5H). ns=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon rank sum test (or Wilcoxon signed-rank test if paired data was available).

FIGS. 6A-6J depict that C. hiranonis is a diet-responsive species with the ability to produce secondary bile acids that inhibit the expansion of potential pathogens in vitro and in vivo. FIG. 6A depicts the Spearman correlations between the abundance of bacteria genera and the levels of bile acids. Only genera that have significant (P<0.05) correlations with bile acids are shown. FIGS. 6B-6E depict the in vitro growth of canine clinical isolates of E. coli (FIGS. 6B and 6C) or C. perfringens (FIGS. 6D and 6E) in the presence of varying concentrations of lithocholic acid or deoxycholic acid (mean±s.d. shown). The in vitro inhibition tests were biologically repeated 2 times. Each point in the graphs represent one replicate well in the assay. FIG. 6F depicts the relative abundance of the OTU corresponding to C. hiranonis (FJ957494.1.1454) in 16S rRNA sequencing data for DR and NDR animals. FIG. 6G depicts the coverage of the bile acid operon (bai) from the C. hiranonis reference (ASM15605v1) with whole genome sequencing reads produced C. hiranonis (teal) and C. perfringens (red) canine clinical isolates. FIG. 6H is a schematic showing experimental design for mouse experiments. FIG. 6I depicts the length of colon at day 8. FIG. 6J depicts E. coli Nissle strain CFUs measured in colon contents at day 8 (mean±s.d. shown for n=5 mice). Experiments were repeated 3 times with similar results. Data shown are from a representative experiment. ns=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon signed rank sum test for relative abundance comparisons or t test for the in vitro culture experiments.

FIGS. 7A-7E depict that the bile acid producer, C. scindens, is associated with diet-induced remission in human pediatric Crohn's disease. Analysis of public data²³ from human pediatric Crohn's disease patients treated with exclusive enteral nutrition (EEN). Relative abundance of reads (mapping ratio) aligning to C. scindens reference (FIG. 7A) or bai operon (FIG. 7B) from 20 patients at pretreatment and 1, 4 and 8 weeks following administration of EEN. Patients that responded to treatment and entered remission (n=10, red) and those that failed therapy (n=10, green) are shown. FIGS. 7C and 7D depict Spearman correlations between log 10-transformed fecal calprotectin levels (FCP) and relative abundance of C. scindens (FIG. 7C) (R=−0.3515 for ‘Responsive’, P=0.0328; R=−0.0267 for ‘Non.Responsive’, P=0.8770) or bai operon (FIG. 7D) (R=−0.3944 for ‘Responsive’, P=0.0157; R=0.0490 for ‘Non.Responsive’, P=0.7766). FIG. 7E is a schematic showing proposed model for diet-induced remission. ns=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon rank sum test for relative abundance comparisons.

FIG. 8 depicts detailed clinical design for canine chronic enteritis study.

FIGS. 9A-9D depict community structures of microbiomes in the dogs with CE and in the healthy dogs. Faith's phylogenetic diversity (FIG. 9A) and Shannon index (FIG. 9B) were compared between the samples from the dogs with CE (day 0) and the samples from healthy dogs. FIG. 9C depicts the ratios of microbiota compositions at a phylum level. FIG. 9D depicts the Unifrac (unweighted) distances within the microbiomes of the dogs with CE or within those of the healthy dogs. ns=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon rank sum test.

FIGS. 10A-10C depict that microbiota community structure changes induced by diet therapy in diet responsive dogs. FIG. 10A depicts Faith's phylogenetic diversity. FIG. 10B depicts Shannon index diversity. FIG. 10C depicts principal coordinate Analysis (PCoA) based on Weighted Unifrac distance of the microbiomes. ns=not significant, *p<0.05, **p<0.01, ***p <0.0001 using Wilcoxon rank sum test.

FIG. 11 depicts that dynamics of microbiome changes at a phylum level for diet responsive dogs (DRs) and non-diet responsive dogs (NDRs).

FIG. 12 depicts principal component analysis based on the abundances of KO (KEGG Orthology) for the samples of day 0 and day 14.

FIG. 13 depicts concentrations of bile acids detected in the fecal samples of diet responsive dogs. NS=not significant, *p<0.05, **p<0.01, ***p<0.0001 using Wilcoxon signed-rank test.

FIG. 14 depicts relative abundance of C. hiranonis in diet responsive dogs calculated from metagenomic data.

DETAILED DESCRIPTION OF THE INVENTION

To date, there remains a need for novel methods and compositions for treating IBD and other intestinal disorders that target gut microbiome and metabolites thereof. The present application relates to method, compositions and food products for improving intestinal health, treating intestinal dysbiosis and/or treating an intestinal disorder in a subject, e.g., a human or a companion animal, which is based, at least in part, on the discovery that intestinal microorganisms that produce bile acids can promote intestinal health and/or is associated with remission from an intestinal disorder after treatment, and that changes of intestinal microorganism population are associated to intestinal health status.

For clarity and not by way of limitation, the detailed description of the presently disclosed subject matter is divided into the following subsections:

1. Definitions;

2. Intestinal bacteria and health assessment tools relating to the same;

3. Pharmaceutical composition;

4. Food products;

5. Treatment methods; and

6 Kits.

1. Definitions

The terms used in this specification generally have their ordinary meanings in the art, within the context of the present disclosure and in the specific context where each term is used. Certain terms are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner in describing the methods and compositions of the present disclosure and how to make and use them.

As used herein, the use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and/or the specification can mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” Still further, the terms “having,” “including,” “containing” and “comprising” are interchangeable and one of skill in the art is cognizant that these terms are open ended terms.

The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, or up to 10%, or up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, e.g., within 5-fold or within 2-fold, of a value.

The term “effective treatment” or “effective amount” of a substance means the treatment or the amount of a substance that is sufficient to effect beneficial or desired results, including clinical results, and, as such, an “effective treatment” or an “effective amount” depends upon the context in which it is being applied. In the context of administering a composition to improving immunity, digestive function and/or decreasing inflammation, an effective amount of a composition described herein is an amount sufficient to improving immunity, digestive function and/or decreasing inflammation, as well as decrease the symptoms and/or reduce the likelihood of a digestive disorder and/or inflammation. An effective treatment described herein is a treatment sufficient to improving immunity, digestive function and/or decreasing inflammation, as well as decrease the symptoms and/or reduce the likelihood of a digestive disorder and/or inflammation. The decrease can be a 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% decrease in severity of symptoms of a digestive disorder or inflammation, or the likelihood of a digestive disorder or inflammation. An effective amount can be administered in one or more administrations. A likelihood of an effective treatment described herein is a probability of a treatment being effective, i.e., sufficient to treat or ameliorate a digestive disorder and/or inflammation, as well as decrease the symptoms.

As used herein, and as well-understood in the art, “treatment” is an approach for obtaining beneficial or desired results, including clinical results. For purposes of this subject matter, beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms, diminishment of extent of a disorder, stabilized (i.e., not worsening) state of a disorder, prevention of a disorder, delay or slowing of the progression of a disorder, and/or amelioration or palliation of a state of a disorder. The decrease can be a 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% decrease in severity of complications or symptoms. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

As used herein, and as well-understood in the art, a “probiotic” is a preparation or composition comprising microorganisms that can provide health benefits when consumed. The microorganisms include, but are not limited to bacteria, fungi, yeasts and archaea. In certain embodiments, the probiotic can modify the microbiome in the GI system to enhance the balance of the microbiome in GI system, e.g., by acting as an inoculum for an increased population of beneficial microbes, and/or by antagonizing growth of deleterious microbes. In certain embodiments, the probiotic is an animal probiotic, e.g., a feline probiotic or a canine probiotic.

As used herein, and as well-understood in the art, a “prebiotic” is a substance or a composition that can induce the growth or activity of one or more beneficial microorganism (e.g., one or more probiotics, e.g., bacteria, fungi, yeasts and archaea). In certain embodiments, the prebiotic can modify the microbiome in the GI system to enhance the balance of the microbiome in GI system. In certain embodiments, the prebiotic is indigestible to an animal. In certain embodiments, the prebiotic can induce the growth or activity of one or more animal probiotics, e.g., a feline probiotic or a canine probiotic.

The term “pet food” or “pet food composition” or “pet food product” or “final pet food product” means a product or composition that is intended for consumption by a companion animal, such as a cat, a dog, a guinea pig, a rabbit, a bird or a horse. For example, but not by way of limitation, the companion animal can be a “domestic” dog, e.g., Canis lupus familiaris. In certain embodiments, the companion animal can be a “domestic” cat such as Felis domesticus. A “pet food” or “pet food composition” or “pet food product” or “final pet food product” includes any food, feed, snack, food supplement, liquid, beverage, treat, toy (chewable and/or consumable toys), meal substitute or meal replacement.

An “individual” or “subject” herein is a vertebrate, such as a human or non-human animal, for example, a mammal. Mammals include, but are not limited to, humans, non-human primates, farm animals, sport animals, rodents and pets. Non-limiting examples of non-human animal subjects include rodents such as mice, rats, hamsters, and guinea pigs; rabbits; dogs; cats; sheep; pigs; goats; cattle; horses; and non-human primates such as apes and monkeys.

As used herein, the term “in vitro” refers to an artificial environment and to processes or reactions that occur within an artificial environment. In vitro environments exemplified, but are not limited to, test tubes and cell cultures.

As used herein, the term “in vivo” refers to the natural environment (e.g., an animal or a cell) and to processes or reactions that occur within a natural environment, such as embryonic development, cell differentiation, neural tube formation, etc. “Pharmaceutical composition” and “pharmaceutical formulation,” as used herein, refer to a composition which is in such form as to permit the biological activity of an active ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to a patient to which the formulation would be administered.

“Pharmaceutically acceptable,” as used herein, e.g., with respect to a “pharmaceutically acceptable excipient,” refers to the property of being nontoxic to a subject. A pharmaceutically acceptable ingredient in a pharmaceutical formulation can be an ingredient other than an active ingredient which is nontoxic. A pharmaceutically acceptable excipient can include a buffer, carrier, stabilizer, and/or preservative.

As used herein, the term “pharmaceutically acceptable salt” refers to any salt of a compound provided herein which retains its biological properties and which is not toxic or otherwise undesirable for pharmaceutical use. Such salts can be derived from a variety of organic and inorganic counter-ions well known in the art. Pharmaceutically acceptable salts further include, by way of example only and without limitation, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium and the like, and when the compound contains a basic functionality, salts of non-toxic organic or inorganic acids.

2. Intestinal Microorganisms and Health Assessment Tools Relating to the Same

The presently disclosed subject matter provides intestinal microorganisms and combinations thereof, which is based, at least in part, on the discovery that intestinal microorganisms that produce bile acids can promote intestinal health and/or is associated with remission from an intestinal disorder after treatment, and that changes of intestinal microorganism population are associated to intestinal health status.

Intestinal Microorganism Capable of Producing a Bile Acid

In certain embodiments, the intestinal microorganism is for use as a medicament. In certain embodiments, the intestinal microorganism is for the treatment of an intestinal disorder in a subject in need thereof.

In certain embodiments, the intestinal microorganism is a bacterium capable of producing a bile acid. In certain embodiments, the bile acid is chenodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, taurochenodeoxycholic acid, taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid or any combination thereof.

In certain embodiments, the bile acid is a primary bile acid. In certain embodiments, the primary bile acid is chenodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, taurochenodeoxycholic acid or any combination thereof.

In certain embodiments, the bile acid is a secondary bile acid. In certain embodiments, the secondary bile acid is taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid or any combination thereof. In certain embodiments, the secondary bile acid is deoxycholic acid and/or lithocholic acid.

In certain embodiments, the bacterium comprises a bile acid-inducible operon (bai operon). In certain embodiments, the bacterium comprises an enzyme having 7-dehydroxylation activity. In certain embodiments, the bai operon comprises a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) homologous or identical to SEQ ID NO: 1 or 3, or any functional fragment thereof. In certain embodiments, the bai operon comprises the nucleotide sequence set forth in SEQ ID NO: 1 or 3. SEQ ID NO: 1 represents an exemplary sequence of C. hiranonis bile acid-inducible operon. SEQ ID NO: 3 represents an exemplary sequence of C. scindens bile acid-inducible operon.

[SEQ ID NO: 1]     1 gcaaattgat tttgattggt atttctttca ttcaaaatat ctcctttcct ttatttagct    61 gtattaaaat ttataaaaaa ttttcattgt taataaaaaa atattctttg ttagtattat   121 agcataattt ataaaaataa tgataatgtt ttaatattga aataataaat atgtaaaaag   181 gttggaaatt tatttaaaaa tgaccagaga taaaaagctc aggtcatttt ttttattatt   241 acaagtaatt tgaaaaaaat atatgaaatg aatggagaaa atataactga gatacatttg   301 ataatgaaaa aaacatttat cgaaattgta aatagactca ttgttataat taataaatat   361 ttattatggc atagttgtta aaattatacc ctaaagaaac gtttcctcaa aaagtgggtt   421 ataaaataaa tgttttttga cgaaagatgt gattttattt gtaccccttt tgtataaaga   481 ttaaacagta tttttgtata aatatattgt atacagtata gagaatgtcg atgtaaaaaa   541 gtatataaaa gtaaataata atcaaaaaaa ctagttttaa ttattaaaaa tgataaaaaa   601 tattaataaa ataaagagtc aaaaatactt gttagttaaa tcacagattt tgtctaagta   661 tagattaggt tttgtatttg aaaaggtcat ctatagtgtt gtaagaaagc gagttattag   721 cacatattgt atctcaaaaa aatgttaaga taatatcaag atagggcgat aaagaaaaaa   781 gcaaattgaa aaagaaaaaa gtaactataa gtttttacaa taaatcaaaa gagaattgat   841 tttaaaagag ggaggcaaaa taccgatatg aatgatgtga aatgtaaata ttttaataaa   901 tttaatacag gaatgtcaga ttttgttact ccaggaaaac agttagaata tgtagcaaaa   961 tgcaagccag atgaaaaagc tatcatatat atagataaag aagacaatgt gagagatatc  1021 acttggaagg aacttcacat agcttcaaat aaactagctt ggcatttaat gaaaaaggga  1081 tttggaaaag gtcaggtagc aatggtatct ttcccaaatg gtatagaaca tatattagca  1141 acattagctg tttggaaaac aggaggttgc tacatgccag tttcttgtaa gataacagat  1201 acagagcttg gtgatatatg cagaataata aaaccaacag tttcttttac agataaagaa  1261 atgccttgta gaacagaaag tataaaaata ggatcagtat tcgatgtttg taaagacgaa  1321 tcagaagaaa tgccagaaga tatagctgca aatccaaata tgatttctcc atctggagga  1381 acaacaggag agcctaagtt cataaaacag aatgtggcaa gtggcttatc tgatgaaatt  1441 ataaaaagct ggtttgaaat gtcaggtatg gaatttgaac aaagacaatt attagtagga  1501 ccacttttcc atggtgctcc tcatacagca gcatttaatg gattatttgt aggaaataca  1561 ttgataatac ctagaaattt aagacctgaa agtatagtta gatatataaa agaatacaaa  1621 atagaattta tacagatgat cccaacatta atgaatagaa taataaaatt agctgatgtt  1681 gataaagaag attttaaatc aataaaagca ctacaccata ctggtggata ttgttctcca  1741 tatttaaaag aaaagtggat cgatataata ggagctgaaa aagttcacga aatgtactct  1801 atgacagagg caatcggtat cacttgtata agaggagatg aatggcttaa acactatgga  1861 agcgtaggac ttccactagg aggaagcaga atatcaataa gagatgaaga aggaaatgaa  1921 ttaggaccac atgaggttgg agaaattcat atgacttcac caagtgcttg ttgcatgaca  1981 gaatacataa accataaacc acttgaaact aaagatggtg gatttagaag tgttggtgat  2041 ttcggttatg tagatgaaga tggatacctt tacttctcag atagaagaag cgacatgctt  2101 gttataggtg gagaaaacgt atttgcgact gaagttgaac cagtactacc agcttatgaa  2161 aaagtagttg atgctgtggt agttggaata cctgatgaag agtggggaag aagattacac  2221 gcaatagtac agaagaaaga agaagtttca gcagaagaat taatcgagta cttaggaaaa  2281 cacttattac catataaagt tccaaagagc tttacatttg ttccttgcat accaagaggt  2341 gacaatggaa aggtaaacag agataagatg ctaaaaggct taatagaaaa aaatctagtt  2401 aataaagttt gctaggatat aaattcagtt aactatctgc accaagtgca gtggaaaata  2461 aatcaaaatt aataaaataa attaataagg taaatttagg aggtctaaaa tgagttacga  2521 cgcacttttt tcaccattta aaatcagagg attagaactt aaaaacagaa tagttctacc  2581 aggtatgaat acaaaaatgg caaaaaataa acatgattta agcgatgata tgatagctta  2641 ccatgttgca agagcaaaag caggttgtgc attaaatata tttgaatgtg ttgcgctatg  2701 tccagcacct catgcatata tgtacatggg attatacaat gacaatcatg tagctcagtt  2761 aaaaaaatta acagatgctg ttcacgaagt tggcggtaaa atggctgttc agttatggca  2821 tggtggtttc agcccacaga tgttctttga taaaacaaat acattagaaa caccagatac  2881 tataacagtt gaacgtattc atgaaatagt taaagagttt ggagaaggtg caagaagagc  2941 tgttgaagct ggattcgatg cagttgaatt ccatgcagca cacagttact tacctcacga  3001 attcctaagt ccaggaatga acaaaagaac tgacgaatat ggtggaaact tcgaaaatcg  3061 ttgcagattc tgcttcgaag tagttgaagc tatacgtgca aatataccag aagatatgcc  3121 attcttcatg agagttgact gcatagatga gttaatggat gaagtaatga cagaagaaga  3181 aatagtagaa ttcataaata gatgtgctga tctaggagta gacgtagctg acttatcaag  3241 aggtaatgct cagtcattcg caacagttta cgaagttcct cctttcaact tacagcacgg  3301 tttcaatata gaaaacatat acaacatcaa aaaacagata aaaataccag taatgggtgt  3361 tggacgtata aacacaggag aaatggctaa ccaggtaata gcagatggaa aatttgactt  3421 agttggtata ggtcgtgctc agttagcaga tcaggattgg gttgctaaag ttagagaagg  3481 taaagaagat ttaatacgtc attgtatagg atgtgaccag ggatgctacg atgcagttat  3541 aaaccctcag atgactcata taacttgtac aagaaaccct cacttatgct tagaatacaa  3601 aggtatgcca aaaactgatg aacctaaaaa agttatgata atcggtggtg gtatggctgg  3661 tatattagca gctgaagtac ttaaaaaacg tggacatgaa ccagttatat tcgaagcttc  3721 tgatcactta gcaggacagt tcgtattagc aggtaaagct ccaatgaaag aagactgggc  3781 agctgcagct aaatgggaag ctgaagaagt agctcgttta ggaatagaag ttagatacaa  3841 tacaaaagtt actccagaat taatagaaga attcgctcca gaccacgttg ttatagctat  3901 aggatctgat tacgtagctc cagctatacc aggtatagat agtgacaaag tttacactca  3961 gtatcaggta ttaaaaggtg aagtagaacc aaaaggacat gtagcagtag ttggttgtgg  4021 attagttggt acagaagttg ctcagtactt agcagctaga ggagctcagg taacagctat  4081 agaaagaaaa ggtgttggta caggtctaag catgcttaga agaatgttca tgaacccaga  4141 attcaaatac tacaaaataa acaaaatgtc tggaactaac atagttggta tagaaccagg  4201 aaaacttcac tacataatga ctaacaagaa aactcaggaa gttactgaag gtgtgttaga  4261 atgtgatgca gcagtaatct gtacaggtat aactgctaga ccaagtgaag atttacagga  4321 aaaatgtaaa gaattaggtg ttccattcaa cgtaataggt gacgcagctg gtgctagaga  4381 tgctagaata gctactcagg aaggttacga agtaggtatg agtatataat ttaaaaatta  4441 tataattata taaattaaaa gttattaaat tacaagaaag aggcgaataa aatgacttta  4501 gaagcaagaa tagaagcatt agaaaaagaa atacagagat taaacgatat agaagctata  4561 aaacagttaa aagctaaata tttccgttgc ctagatggaa aattatggga tgaattagaa  4621 actactcttt ctcctaacat agaaacttct tactctgatg gaaaattagt attccacagc  4681 ccaaaagaag taactgaata tttagcagca gcaatgccta aagaagaaat aagtatgcac  4741 atgggacata ctccagaaat aactatagac agcgaaaata ctgctacagg aagatggtac  4801 ttagaagata acctaatatt cacagacgga aaatacaaaa acgttggaat aaacggtgga  4861 gcattctaca cagataaata tgaaaaaata gacggacagt ggtacataaa agaaactgga  4921 tatgttcgta tatttgaaga acatttcatg agagatccaa aaatacatat aactagcaac  4981 atgcataaag aaaaataata actgattgct aataaacaag atataaacag ggggctggta  5041 aacagccagc cctctgaaaa ataaactaaa aaactataat cttttaaaat cttaattaaa  5101 gtagaaggag ataagacaat gaacttagta caggacaaaa tagttataat aacaggtgga  5161 acaagtggta taggtctttg cgcagcaaaa atattcatgg ataacggtgc aacagtttct  5221 atattcggaa aaactcagga agaagtagat gctgctaaag cagaattaaa agaaactcac  5281 ccagataaag aagtattagg atttgctcca gatttaacta atagagatga agttatggct  5341 gcagttggtg cagtagctga aaaatacgga agattagacg ttatgataaa caatgctggt  5401 gttactagct caaacgtatt ctcaagagtt agcccagaag aattcacata tttaatggat  5461 ataaacgtta caggtgtatt ccatggtgct tgggctgctt accactgcct gaaaggtgaa  5521 aagaagatta taataaatac tgcttcagta acaggaatac acggatcatt atcaggagtt  5581 ggatacccaa caagtaaatc agctgttgta ggattcactc aggctcttgg tagagaaata  5641 atacgtaaaa acataagagt tgttggtgtt gcaccaggtg ttgttaacac tccaatggtt  5701 ggtaatatac cagatgaaat attagatgga tacctaagct cattcccaat gaagagaatg  5761 ttagaaccag aagaaatagc taacacttac ttattcttag cttctgactt agctagtggt  5821 ataacagcta caactgtaag cgttgacggt gcttatagac catcataaga tttactttaa  5881 tttaaaactg taattagata gataatacga cgattaatat aaaaaatgtt ctttaaaaga  5941 aaaggagaaa taaaatggct ggattaaaag attttcctaa atttggtgca ctttctggat  6001 taaaaatatt agatagtgga tctaacatag ctggacctct aggtggtgga cttttagcag  6061 aatgtggtgc tacagttata cacttcgaag gacctaaaaa acctgacaac cagagaggtt  6121 ggtatggata ccctcagaac cacagaaacc agttatcaat ggttgctgat ataaaatctg  6181 aagaaggtag aaaaatattc ttagacttaa taaaatgggc tgacatatgg gttgaatcat  6241 caaaaggtgg acagtacgac agactaagtc tttctgatga agttatatgg tcagtaaacc  6301 ctaaaatagc tatagttcac gtttctggat acggacaggt tggagatcca tcatacgtaa  6361 caaaagcttc ttatgatgct gttggacagg cattcagtgg atacatgtca ttaaatggtg  6421 ttaatgaagc attaaaaata aatccttacc taagtgactt cgtatgtgtt cttactactt  6481 gctgggcaat gttagcatgc tacgtaagta ctcagttaac tggaaaagga gaatctgtag  6541 acgttgctca gtacgaagca ttagctcgta taatggacgg acgtatgata cagtacgcta  6601 ctgatggtgt aagtgttcca aaaactggta acaaagatgc tcaggcagct ctattcagct  6661 tctatacttg taaagatgga agaactatat tcataggtat gactggtgct gaagtatgta  6721 agagaggatt ccctgtaata gggcttccag ttcctggtac aggtgaccct gacttcccag  6781 aaggattcac aggatggatg ataaatactc cagttggaca gagaatggaa aaagctatgg  6841 aagcattcgt tgctgaaaga actatgccag aagttgaaaa agctatgata gatgctcaga  6901 taccatgcca gagagtttat gatcttgaag actgcttaaa cgaccctcac tggaatgctc  6961 gtggaactat aatggaatgg gatgacccaa tgatgggaca cataaaaggt cttggattaa  7021 taaacaaatt caaaaacaac ccttctgaaa tatggagagg tgctccatta ttcggtatgg  7081 acaacagaga cataattaga gaccttggat attctgagga ggaagttaac gatttatacg  7141 ctaaaggtat tgtaaacgaa ttcgaccttg aaacaactat aaaacgttac aaacttgatc  7201 aggttatacc tcacatggct aaaaaagata aataagaaac gtattaaata ataaaatata  7261 aatgtcgagc ctgccagaat gagaattttg acaggcttga tattataacg aaatgttata  7321 aaaaaaacaa aataaaaatt gcttaaattt tatacaagga gaattgaaat gacagcaaca  7381 aacgcaaact ataaaaaagg ctttatccca tttgctatag cagcgttact agtaggtctt  7441 ataggtggtt tcacagccgt tctagcacct gcattcgtag cagatatggg tcttaacgat  7501 aacaatacta catggatagc actagcgctt gcaatgtcta cagctgcatg tgctccaata  7561 cttggtaaat taggtgacgt acttggacgt cgtaaaactt tattattagg aatcatagta  7621 ttcacaatag gtaacgtatt aacagcaata gcatcttcat taatattcat gctaggtgca  7681 agatttatag ttggggttgg tacagcggct atagctccag ttataatggc ttacatagtt  7741 acagaatatc caccagaaga aactggtaag ggattcgctc tttatatgtt aatatcaagt  7801 gctgcagttg ttgttggtcc aacttgtggt ggattaataa tgcaggcatt tggatggaga  7861 atgatgatgt gggtttgtgt tgccctttgt gtagtaacat tcttcatatg ttcagtaatg  7921 attaagaaaa cagactttga aaagaaaagt cttgataact tcgataaaaa aggtgcagta  7981 tgcgtactaa tattcttcag tttagtatta tgtataccat catttggaca gaatataggt  8041 tggacatcag cgccattcct aggtgttaca gcagtagctt tagtaacatt attcttatta  8101 ataaaagctg aaagcagtgc agaaaaccca atattaagtg gtaaatttat gaaacgtaaa  8161 gaattcatat taccagtatt aatattattc cttactcagg gattaatgca ggctaacatg  8221 actaacgtaa tattattcgt tagagctact cagccagaaa atacaataat atcaagtttc  8281 gcaatatcaa tcctttacat aggtatgtct ttaggttcag tattcatagg acctatggca  8341 gataaaaaag aaccaaaaac tgtacttaca ggatcacttc tattcactgg tataggttgt  8401 gcaatgatgt acttcttcac agaaactgca ccattcgcaa tgttagctgg atctctagga  8461 atgttaggta taggacttgg aggaaatgct acaatactaa tgaaagtttc attatctgga  8521 ttatctcagg cagaagctgg atcaggaaca ggaacatacg gattattcag agatatatca  8581 gctccatttg gtgttgcggt attcgtacca ctatttgcaa acacagttac aacaagaatg  8641 gctggagtaa tggctaacgg aactgcagaa gctgctgcta aatcattagc atctgtttct  8701 tctatacata cattagcatt agttgaagta tgctgtgtaa tattagcaat agttgcagtt  8761 agaatgctac caaaaataca caataaataa tttaaaaata ataacagagt tgaaaaaaca  8821 ctcaattaaa agaggggcct tgagcccctt ttttagtgta aaaatgacaa aatactatca  8881 atttatataa atgataatta aactcgtcaa ccaaagaaat attcacaaag tagataataa  8941 tagatattca aaaagtgata tattattagg caaaaagtgc aagaaattag cgagtattcg  9001 acaacttttt gtccaatggt agaaaagaat atttgttatc ataaatatag acaaagggct  9061 ttgaccaaaa ctaaggaaaa agtttgcata atataaaaaa taaaataaaa taaaaaaata  9121 aaaataaaat aaaagcgaaa ggaaaaaaca acatcatgga tatgaaaaat tctaaactat  9181 tctcaccttt aacaatagga tcattaacat taaacaacag agttggtatg gcaccaatga  9241 gtatggacta cgaagctgct gacggaacag ttccaaaaag attagcagat atatttgttc  9301 gtagagctga aggtggaaca ggatatgtaa caatagacgc ggtaacaata gatagtaaat  9361 ataaatatat gggtaataca actgctttag attctgatga tttagttact cagttcaaag  9421 aatttgcaac aagagttaga gaagcaggaa gcacattaat acctcaggtt atacatccag  9481 gaccagaatc aatatgtgga tacagacaca tagcaccact tggaccatca gttaatacaa  9541 atgctaactg ccacgtgagc cgtgctataa gtgtagatga aatacatgaa ataataaaac  9601 agtttggaca ggctgctaga agagttgaag aagcaggatg cggtggtata ggattacact  9661 gtgcacatgc ttacatgcta ccaggttcat tcttatctcc attaagaaac aaaagaatgg  9721 atgaatacgg cggatgtcta gataacagag caagattcgt aatagaaatg atagaagaag  9781 ttcgtagaaa tgtaagtcct gatttcccaa taatgcttag aatatctggg gatgaaagaa  9841 tgataggagg aaactcttta gaagatatgt tatacttagc tccaaaattt gttgaagctg  9901 gtgtaaatat gtttgaagtt tctggaggta ctcagtacga aggattagaa cacataatac  9961 caagtcagaa caaaagcata ggtgtaaacg tacacgaagc atctgaaatc aaaaaagttg 10021 tagatgttcc agtttacgct gttggtaaaa taaatgacat aagatacgct gctgaaatag 10081 ttgaaagagg actagttgat ggggtatcaa taggtagacc attattagca gatccagact 10141 tatgtaataa agcaaaagaa aacttatttg atgaaataac tccatgtgca agctgtggag 10201 gaagctgtat aagccgtact gcagatagac ctcagtgtcg ttgccatata aacccaagag 10261 ttggattcga atatgattat ccagaagttc cagctgaaaa atctaaaaaa gttctagttg 10321 taggtgctgg acctggtggt atgatggcag cagttacagc agctgaaaga ggacatgatg 10381 taacactttg ggaagctgac actcagatag gtggacagat aaacttagca gtagtagctc 10441 caggtaaaca ggaaatgact aaatggttat ctcacttaaa ctacagagct aaaaaagctg 10501 gagttaaaat ggtattagga aaagaagcta cagtagaaaa cataaaagaa tttgctccag 10561 aagcagttat agttgcaaca ggtgctagac cattagttcc accaataaaa ggaactcagg 10621 actacccagt tcttacagct catgacttct taagaggaaa attcgttata ccaaaaggaa 10681 aagtttgtgt actaggtgga ggagctgttg cttgtgaaac tgcagaaaca gtattagaaa 10741 acgctagacc aaacgcattc actagaggat ttgatgctag tatcggtgat gtagatgtta 10801 cattagtaga aatgttacca cagttattaa caggagtatg tgctccaaat agaactccat 10861 taataagaaa acttaaaaac aaaggtgttc atataaatgt aaatactaaa atattagaag 10921 taactgacca cgacgttaaa gttcagagag ctgacggtgc agaagaatgg ttaaaaggat 10981 tcgactacat actattcgga cttggttcta gaaactacga tccaatatct gaacagataa 11041 aagaattcgt tccagaagta cacgttgttg gggatgctaa gagagctaga caggcaagct 11101 ttgcaatgtg ggaagctttc gaagcagcat acagcttata a [SEQ ID NO: 3]     1 aaaagatatt aagcattaag aaaatgcaca aaaaatcagc gtgtgagagg gagggcaagg    61 agttgaagcg tgactttttt aacaagttta atttggggac atcgaacttt gtcacgccgg   121 gaaaacagtt ggaatacgtt tcggaatgca agccagattc tactgcggtc atttgcttag   181 ataaagaaca gaactgttcc gttattactt ggcatcagct gcacgtctat tccagccagc   241 tggcatggta ccttatagaa aatgagattg gcccggggtc gatcgtactt acaatgtttc   301 cgaacagcat cgagcacatt attgcggtat ttgcaatctg gaaggcgggc gcctgctata   361 tgcccatgtc ctataaggcg gcggaatccg agatcaggga ggcctgcgat accatccacc   421 cgaatgcggc ttttgcggaa tgcaagattc caggattaaa attctgcctt agcgcagacg   481 agatatatga ggcgatggaa ggaagatcca aggagatgcc ttcggaccgt ctggccaatc   541 cgaacatgat atccttatca ggcggaacca gcggaaagat gaagttcatc cgtcagaacc   601 ttccatgcgg gctggacgat gagacgatca gaagctggtc tttgatgtct ggaatgggat   661 ttgagcagcg ccagctgctg gtaggcccgc tgtttcatgg cgcgcctcac tccgcggcgt   721 ttaatggact gttcatgggc aacaccctgg tactgaccag gaacctttgc ccgggaaata   781 tcctgaacat gattaagaaa tataagattg aatttataca gatggtgccg accctgatga   841 accggcttgc caaactggag ggagtcggaa aagaagactt tgcatccctg aaggcgctgt   901 gccatacagg gggcgtctgt tctccctggc ttaagcagat ctggatcgac ctgctggggc   961 ctgaaaagat ctatgagatg tattccatga cggaatgcat cggccttacc tgcatccggg  1021 gagacgagtg ggtgaagcat ccgggaagca tcggacggcc agtgggcgat agcaaggtgt  1081 ctatccggga tgagaatggc aaggaagttg cgccttttga gattggcgag atctatatga  1141 cagcgccggc ctcctatctg gttaccgagt acatcaattg ggaaccgctg gaagtgaaag  1201 agggaggctt ccgaagcgta ggggatatcg gctacgtgga tgagcagggc tatctgtact  1261 tttctgaccg gcgcagcgac atgctggtat caggcggaga aaacgtgttc gccaccgaag  1321 tcgagacggc gcttttgaga tataaggata tcctggacgc tgtagtggta gggataccgg  1381 atgaagatct ggggcgaagg ctccatgcgg tcattgagac agggaaagag ataccggcag  1441 aggaactgaa aacattcctg agaaagtatc tgactccata taagatacca aagacgttcg  1501 agttcgtaag gagcatacga aggggagaca atggaaaggc cgacaggaag cggatcctgg  1561 aagattgtat tgcccgcggg ggatgattct ataaatgcaa agaaaacaaa ttatataaag  1621 gaggagtaac aaaatgagtt acgaagcact tttttcacca ttcaaggtca gaggactgga  1681 acttaaaaac cgtatcgtcc tgcctggaat gaacaccaag atggcaaaga acaagcacga  1741 cataggcgag gatatgatag cctaccatgt tgccagggca aaagcgggat gcgcgttaaa  1801 tatatttgaa tgcgtagcat tatgtccggc gcctcacgct tatatgtata tggggcttta  1861 tacggaccat catgtagaac agcttaagaa attgacggat gcagtccatg aagcaggcgg  1921 caagatgggc atccagctgt ggcatggagg attcagcccg cagatgttct ttgacgagac  1981 caacaccctg gaaactccgg acactcttac ggtagagagg attcatgaga tcgtagaaga  2041 attcggacgc ggcgcaagga tggctgttca ggctggattt gacgcagtag aattccatgc  2101 ggctcacagt tatctgcctc acgagttctt aagccctgga atgaacaaac gtacggatga  2161 gtacggcgga agttttgaga accgctgcag attctgttat gaagtcgttc aggcaatccg  2221 ttccaatatc ccggatgaca tgccattctt tatgcgtgca gactgcatcg acgaattaat  2281 ggaacagacc atgacagagg aagagatcgt tacatttatc aataagtgcg cagaacttgg  2341 cgtggatgtg gcagaccttt cccgtggaaa cgcgacttca ttcgcaaccg tatatgaagt  2401 tccgccattc aacctggctc atggcttcaa catagagaat atttacaaca tcaaaaagca  2461 gatcaatatc ccggttatgg gagttggccg tatcaataca ggagagatgg caaacaaggt  2521 cattgaagaa ggcaagtttg acctggtagg catcggacgc gcccagcttg cagatccaaa  2581 ctggatcacc aaagtaagag aaggcaaaga agacctgatc cgccactgta tcggatgtga  2641 ccagggatgc tatgacgcag tcatcaatcc aaagatgaag catatcacct gcacccacaa  2701 tccaggattg tgcttagagt atcagggaat gccaaagaca gacgctccta agaaagtcat  2761 gatcgtagga ggcggaatgg caggcatgat cgctgcggaa gtattaaaga ccagaggcca  2821 taacccggta atcttcgagg catccgacaa gcttgcagga cagttcaggc tggcaggcgt  2881 agcgccgatg aagcaggatt gggcagatgt tgcagaatgg gaagcaaaag aagtagagcg  2941 ccttggaatc gaagtacgtc tgaataccga agtgactgca gagaccatca aggaattcaa  3001 tccggataat gtcatcatcg cagtaggctc tacctatgcg ctgcctgaga ttccgggaat  3061 cgacagccca agcgtatact cccagtatca ggtactgaaa ggggaagtaa atccgacagg  3121 ccgtgtagcc gttatcggat gcggactggt tggtacggaa gtcgcagaac ttctggcatc  3181 cagaggcgca caggtaatcg cgatcgagag gaagggcgta ggtaccggcc ttagcatgct  3241 tcgcagaatg ttcatgaacc cggaattcaa atattacaag atcgccaaga tgtccggaac  3301 aaatgtcacc gctttagagc agggcaaggt tcactacatc atgacagaca agaagaccaa  3361 agaagtgacg cagggagtcc tggaatgcga cgctaccgtt atctgtacag gaattaccgc  3421 acgtccaagc gatgggctta aggcaagatg cgaagaactt ggaatcccgg ttgaggtgat  3481 cggagacgct gctggcgcaa gagactgcac gatcgcgaca cgcgaaggct atgacgcagg  3541 aatggcaatc tagaaaatca gaacttatca atcttacata tagaaaggat gatacatatg  3601 acattagaag agagagttga agcattagaa aaagaattgc aggagatgaa ggatattgag  3661 gcaatcaagg aactgaaagg aaagtatttc cgctgcctgg acggaaagat gtgggatgag  3721 ctggagacca ccctgtcacc aaatatcgta acctcttatt ccaacgggaa actggtattc  3781 catagcccga aggaagttac cgattactta aagagctcga tgccaaaaga agagatcagc  3841 atgcatatgg gccacacgcc ggagatcacc attgacagcg agactacggc tacgggcaga  3901 tggtatctgg aagatagact gatctttacg gacggtaagt acaaagacgt aggaatcaat  3961 ggcggcgcgt tctatacaga caaatatgag aagatagacg gccagtggta catccttgaa  4021 accggctatg tacgaatcta tgaagaacat ttcatgcgtg atccaaagat ccatatcacg  4081 atgaacatgc acaaataaga atattgtaaa agaaaggcag gagtaagagt atgaatctcg  4141 tacaagacaa agttacgatc atcacaggcg gcacaagagg tattggattc gccgctgcca  4201 aaatatttat cgacaatggc gcaaaagtat ccatcttcgg agagacgcag gaagaagtag  4261 atacagcgct tgcacagtta aaagaacttt atccggaaga agaggttctg ggattcgcgc  4321 cggatcttac atccagagac gcagttatgg cagcggtagg ccaggtagca cagaaatatg  4381 gcagactgga tgtcatgatc aacaatgcag gaattaccag caacaacgta ttctccagag  4441 tgtctgaaga agagttcaag catattatgg acatcaacgt aacaggcgta ttcaacggcg  4501 catggtgcgc ataccagtgc atgaaggatg ccaaaaaggg cgttatcatc aacacggcat  4561 ccgttacagg catcttcgga tcactctcag gcgtaggata tccggccagc aaggcaagcg  4621 tgatcggact cacccatgga cttggaagag agatcatccg caagaatatc cgtgtagtag  4681 gagtggctcc tggagttgtg aacacggata tgaccaatgg caatcctccg gagatcatgg  4741 aaggatatct gaaggcgctt ccgatgaaga gaatgcttga gccggaagag atcgctaatg  4801 tatacctgtt cctggcatct gacttggcaa gcggcattac ggctactacg gtcagcgtag  4861 acggggctta cagaccataa ttttaatttt tactaagtag aatatgtgat atagaaaagg  4921 agatataaaa acatggctgg aataaaagat tttccaaaat tcggagctct tgcagggctt  4981 aagatacttg acagcggatc taacatcgcc ggacctttag gcggaggcct tctggcagaa  5041 tgcggagcaa cggtcatcca ttttgaagga ccaaagaaac ctgataacca gagaggatgg  5101 tacggctatc cacagaatca ccgtaatcag ctgtctatgg tagcagacat caaatctgaa  5161 gaaggaagaa agatcttcct tgatctgatc aaatgggcag atatctgggt agagtcatcc  5221 aaaggcggac agtatgacag gctgggactt tccgatgaag tcatctggga agtaaatcct  5281 aagattgcca tcgtgcacgt atccggatat ggacagacag gagacccgtc ttacgttaca  5341 cgtgcatcct atgacgcagt aggccaggca ttcagcggct atatgtcact gaacggaaca  5401 acggaagcgc tgaagatcaa tccttatctg agcgatttcg tatgcggact taccacatgc  5461 tgggctatgc ttgcctgcta tgtaagcacc attcttaccg gaaaaggcga atctgttgac  5521 gttgcacagt acgaagcgct ggcacgtatc atggacggac gtatgatcca gtacgctaca  5581 gacggcgtga agatgccaag aaccggcaat aaggatgcgc aggctgccct gttcagcttc  5641 tacacctgta aagacggacg tacgatcttt atcggaatga ctggcgcgga agtatgtaag  5701 agaggcttcc cgatcatcgg acttccggta cctggaaccg gagacccgga cttcccggaa  5761 ggcttcacag gctggatgat ctatactcct gtaggacaga gaatggaaaa ggctatggag  5821 aagtatgtat ctgagcatac gatggaagaa gtagaggctg agatgcaggc acaccagatt  5881 ccatgccaga gagtatacga gctggaagac tgcctgaacg atcctcactg gaaagcacgt  5941 ggaactatta cggagtggga tgacccgatg atgggacata tcacaggcct tggactgatc  6001 aacaagttca agagaaatcc ttccgaaatc tggagaggcg ctccgctgtt cggtatggat  6061 aaccgcgata tcctgaaaga cctgggatat gacgatgcaa agatcgatga actctatgag  6121 cagggcatcg tcaatgaatt cgaccttgac actactatca aacgctatag actggatgaa  6181 gtaattccac atatgagaaa gaaagaggag taagagtatg agcaccgtag ccaatccaaa  6241 ttataagaaa ggttttgtcc cctttgcaat tgcagcactc ctggtgagcc tgatcggcgg  6301 ttttaccgcc gttctcggcc cggccttcgt ggcggaccag gggattgact ataataatac  6361 cacatggatt tccctggcgc tggcgatgtc ttccgccgca tgcgctccaa tccttggaaa  6421 actgggagac gtgctaggac gcaggacgac gctgcttctg ggtattgtga tctttgcggc  6481 cggcaatgtg ctgacagccg tagccacgtc cctgatattc atgctggcag cccgttttat  6541 cgtaggtatc ggaacagcag cgatctcacc gatcgttatg gcctatatcg taaccgagta  6601 tccgcaggag gagacaggaa aggcctttgg cctgtatatg ctgatctcca gcggcgccgt  6661 cgtggtagga cctacctgtg gcggcctgat catgaatgcg gctggctgga gagtcatgat  6721 gtgggtatgc gtcgctctgt gcgtcgttgt attcctgatc tgcacattct ccatcaagaa  6781 gactgcattt gagaagaaga gcatggcagg atttgacaag ccgggcgcag ccctggtagt  6841 cgtattcttc agtttgttcc tgtgcatccc atccttcgga cagaatatcg gatggtcttc  6901 cacagcattt atcgcagcag cggcagtagc gctggtagca cttttcatcc tggtaatggt  6961 agaaaagaaa gcgaagagtc cgatcatgaa cggcaagttt atggcacgca aggaattcgt  7021 gcttccagta ttgatcctgt tccttacaca gggacttatg atggcaaata tgaccaatgt  7081 catcgtgttc gtgcgctata cgcagccgga caatgtcatt atatcaagtt ttgcgatctc  7141 catcatgtac ataggaatgt ccttaggctc cgttatcatt ggacctgttg cagataagaa  7201 agagccaaag acggttctga cattctctct ggtactgaca gccatcggct gtgcgctgat  7261 gtatctgttc aaggcagatt cctccgtcgc tatctttgcg gcatccttgg gaatccttgg  7321 atttggcctt ggaggaaatg caaccatctt catgaaggta gcgctttccg gcctgtccag  7381 cgaagtagct ggctctggta ctggaaccta tggcctgttc agagatatct cggcaccatt  7441 cggcgtggca gtgttcgtgc ctatgtttgc caacggcgta acagcgaata ttgcgaaata  7501 cgcgtcaggc ggcatggaag aaggcgccgc tacggtaaaa gcagccatct catccatcca  7561 gacgctgaca ctggttgaac ttggatgtat cgttgtggga atcatccttg tgagaatgct  7621 gccaagaatc tatcagaaga aagaggcata aataagttaa gaaaagaggt aattataaat  7681 ggatatgaaa cattccagat tattttcgcc gcttcagatc ggatccctga cactgtctaa  7741 ccgtgtcggc atggctccca tgagcatgga ctatgaagca gcagacggaa ctgtgcccaa  7801 gaggctggcg gacgtatttg tccgccgcgc cgagggaggc acaggctacg tcatgatcga  7861 cgcggtgacg atagacagca agtatcctta tatgggaaat acaacggccc ttgaccgtga  7921 tgaactggtt ccccagttta aggaatttgc tgacagagta aaagaagcag gcagcacgct  7981 ggtgccgcag atcattcatc cgggtccgga atccgtatgc ggctaccggc atatcgctcc  8041 gcttggacct tctgccaaca ccaatgcaaa ctgccacgtg agcagatcga tcagcataga  8101 tgagatccat gacatcatta agcagttcgg ccaggcggca cgccgcgccg aagaagcagg  8161 atgcggggca atctccctgc actgcgcgca tgcgtatatg ctgccaggat ccttcctgtc  8221 accgcttcgc aacaagcgca tggatgaata tggcggaagc cttgacaacc gtgcccgttt  8281 cgtgatcgag atgattgagg aggcccgcag gaatgtgagt cctgatttcc cgatcttcct  8341 tcgtatctcc ggagacgaga gaatggtagg aggcaacagc cttgaagata tgctctacct  8401 ggcaccgaag ttcgaggctg ccggcgtaag catgctggaa gtatccggcg gaacccagta  8461 tgaaggcctg gaacatatca ttccttgcca gaataagagc aggggcgtca atgtatatga  8521 agcttctgag atcaagaaag tagtgggcat cccggtatac gcagtaggaa agatcaacga  8581 tatacgctat gcggcagaga tcgtagaacg cggcctggta gacggcgtgg ctatgggacg  8641 tccgcttctg gcagatccgg acctttgcaa gaaggcagtg gaaggccagt ttgacgagat  8701 cactccatgc gcaagctgcg gcggaagctg catcagccgt tctgaggcag cgcctgagtg  8761 ccattgccat attaatccaa ggcttggccg ggagtatgaa ttcccggatg tgcctgccga  8821 gaagtccaag aaggtactgg ttatcggcgc aggccctgga ggaatgatgg ctgccgtgac  8881 agctgcggaa cgcggccatg atgttacggt atgggaggct gacgacaaga tcggcggcca  8941 gctgaacctg gcagtagtgg ctcctggcaa gcaggagatg acccagtgga tggtacatct  9001 gaactatcgc gcgaagaaag caggcgtgaa gtttgaattc aataaagaag cgacggcaga  9061 agatgtcaag gcgctggcgc cggaagcagt gatcgttgct acaggcgcga agccgctggt  9121 tcctccgatt aaaggaacac aggattatcc ggtgcttact gcccatgatt tccttcgcgg  9181 caagttcgtg attccgaagg gacgcgtctg cgtgctggga ggaggcgcgg ttgcctgcga  9241 gactgccgag acagccctgg agaatgcacg tccgaattct tataccagag gatacgatgc  9301 aagcatcgga gatatcgatg tcacgcttgt ggagatgctt ccgcagctcc ttaccggcgt  9361 atgcgcgccg aaccgcgagc ctttgatccg caagttaaag agcaagggcg tacacatcaa  9421 cgtcaatacc aagatcatgg aagtaacaga ccatgaagta aaggttcaga gacaggatgg  9481 aacgcaggaa tggctggaag gatttgacta tgtcctcttt ggccttggtt ccagaaatta  9541 cgatccgctt tcagagaccc tcaaggaatt cgttccggaa gtacatgtca tcggcgatgc  9601 cgtaagggcg cgccaggcaa gctacgcaat gtgggaagga tttgagaagg catacagcct  9661 gtaaaagcgg tttgagtaaa aggaggctta agaaatggca gtgaaggcaa tctcaggctg  9721 cgacaaggat caggaactga tca

In certain embodiments, the bacterium is transformed with a vector comprising a bile acid-inducible operon (bai operon). In certain embodiments, the bacterium stably expresses an enzyme having 7-dehydroxylation activity. In certain embodiments, the enzyme is a bile-acid 7-dehydroxylase. In certain embodiments, the enzyme is selected from the group consisting of a bile-acid 7-dehydroxylase, bile-acid 7-alpha-dehydroxylase, 7-alpha-dehydratase, bile acid CoA ligase, 3 alpha-HSDH, CoA transferase, 3-dehydro-4-7-alpha-oxidoreductase, 3-dehydro-4-7-beta-oxidoreductase, CA/CDCA transporter, 7-beta-dehydratase and AraC/XyIS. In certain embodiments, the enzyme comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) homologous or identical to the amino acid of a bile-acid 7-dehydroxylase, bile-acid 7-alpha-dehydroxylase, 7-alpha-dehydratase, bile acid CoA ligase, 3 alpha-HSDH, CoA transferase, 3-dehydro-4-7-alpha-oxidoreductase, 3-dehydro-4-7-beta-oxidoreductase, CA/CDCA transporter, 7-beta-dehydratase or AraC/XyIS.

In certain embodiments, the bacterium is selected from the group consisting of Ruminococcus, Alloprevotella, Allisonella, Anaerostipes, Anaerobiospirillum, Bacteroides, Blautia, Clostridium sensu stricto 1, Collinsella, Coprococcus 1, Corynebacterium 1, Campylobacter, Enterococcus, Erysipelatoclostridium, Escherichia-Shigella, Faecalitalea, Fusobacterium, Clostridium, Helicobacter, Intestinibacter, Lachnoclostridium, Lactobacillus, Megasphaera, Methanobrevibacter, Parabacteroides, Porphyromonas, Phascolarctobacterium, Peptoclostridium, Prevotellaceae UCG-001, Pseudocitrobacter, Ruminiclostridium 9, Sarcina, Streptococcus, Succinivibrio, Treponema 2, Turicibacter, Tyzzerella, Tyzzerella 4 and any combination thereof. In certain embodiments, the bacterium is selected from the genus of Clostridium.

In certain embodiments, the intestinal microorganism comprises a 16s rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) homologous or identical to SEQ ID NO: 2. In certain embodiments, the intestinal microorganism comprises a 16s rRNA comprising the nucleotide sequence set forth in SEQ ID NO: 2 or 4. SEQ ID NO: 2 represents an exemplary sequence of 16S rRNA gene in C. hiranonis. SEQ ID NO: 4 represents an exemplary sequence of 16S rRNA gene in C. scindens.

[SEQ ID NO: 2]    1 acatgcaagt cgagcgattc tcttcggaga agagcggcgg acgggtgagt aacgcgtggg   61 taacctgccc tgtacacacg gataacatac cgaaaggtat gctaatacgg gataatatat  121 aagagtcgca tgacttttat atcaaagatt tttcggtaca ggatggaccc gcgtctgatt  181 agcttgttgg cggggtaacg gcccaccaag gcgacgatca gtagccgacc tgagagggtg  241 atcggccaca ttggaactga gacacggtcc aaactcctac gggaggcagc agtggggaat  301 attgcacaat gggcgcaagc ctgatgcagc aacgccgcgt gagcgatgaa ggccttcggg  361 tcgtaaagct ctgtcctcaa ggaagataat gacggtactt gaggaggaag ccccggctaa  421 ctacgtgcca gcagccgcgg taatacgtag ggggctagcg ttatccggat ttactgggcg  481 taaagggtgc gtaggcggtc tttcaagtca ggagttaaag gctacggctc aaccgtagta  541 agctcctgat actgtctgac ttgagtgcag gagaggaaag cggaattccc agtgtagcgg  601 tgaaatgcgt agatattggg aggaacacca gtagcgaagg cggctttctg gactgtaact  661 gacgctgagg cacgaaagcg tggggagcaa acaggattag ataccctggt agtccacgct  721 gtaaacgatg agtactagtt gtcggaggtt accccttcgg tgccgcagct aacgcattaa  781 gtactccgcc tggggagtac gcacgcaagt gtgaaactca aaggaattga cggggacccg  841 cacaagtagc ggagcatgtg gtttaattcg aagcaacgcg aagaacctta cctaggcttg  901 acatccttct gaccgaggac taatctcctc tttccctccg gggacagaag tgacaggtgg  961 tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1021 cccttgtctt tagttgccat cattaagttg ggcactctag agagactgcc agggataacc 1081 tggaggaagg tggggatgac gtcaaatcat catgcccctt atgcctaggg ctacacacgt 1141 gctacaatgg gtggtacaga gggcagccaa gccgtgaggt ggagcaaatc ccttaaagcc 1201 attctcagtt cggattgtag gctgaaactc gcctacatga agctggagtt actagtaatc 1261 gcagatcaga atgctgcggt gaatgcgttc ccgggtcttg tacacaccgc ccgtcacacc 1321 atgggagttg gagacacccg aagccgacta tctaaccttt tgggagaagt cgtccccctc 1381 gaatcaatac ccc [SEQ ID NO: 4]    1 gagagtttga tcctggctca ggatgaacgc tggcggcgtg cctaacacat gcaagtcgaa   61 cgaagcgctt ccgctagatt ttcttcggag atgaaggcgg ctgcgactga gtggcggacg  121 ggtgagtaac gcgtgggcaa cctgccttgc actgggggat aacagccaga aatggctgct  181 aataccgcat aagaccgaag cgccgcatgg cgcagcggcc aaagccccgg cggtgcaaga  241 tgggcccgcg tctgattagg tagttggcgg ggtaacggcc caccaagccg acgatcagta  301 gccgacctga gagggtgacc ggccacattg ggactgagac acggcccaga ctcctacggg  361 aggcagcagt ggggaatatt gcacaatggg ggaaaccctg atgcagcgac gccgcgtgaa  421 ggatgaagta tttcggtatg taaacttcta tcagcaggga agaagatgac ggtacctgac  481 taagaagccc cggctaacta cgtgccagca gccgcggtaa tacgtagggg gcaagcgtta  541 tccggattta ctgggtgtaa agggagcgta gacggcgatg caagccagat gtgaaagccc  601 ggggctcaac cccgggactg catttggaac tgcgtggctg gagtgtcgga gaggcaggcg  661 gaattcctag tgtagcggtg aaatgcgtag atattaggag gaacaccagt ggcgaaggcg  721 gcctgctgga cgatgactga cgttgaggct cgaaagcgtg gggagcaaac aggattagat  781 accctggtag tccacgccgt aaacgatgac tactaggtgt cgggtggcaa ggccattcgg  841 tgccgcagca aacgcaataa gtagtccacc tggggagtac gttcgcaaga atgaaactca  901 aaggaattga cggggacccg cacaagcggt ggagcatgtg gtttaattcg aagcaacgcg  961 aagaacctta cctgatcttg acatcccgat gccaaagcgc gtaacgcgct ctttcttcgg 1021 aacatcggtg acaggtggtg catggttgtc gtcagctcgt gtcgtgaggt gttgggttaa 1081 gtcccgcaac gagcgcaacc cctatcttca gtagccagca tttcggatgg gcactctgga 1141 gagactgcca gggacaacct ggaggaaggt ggggatgacg tcaaatcatc atgcccctta 1201 tgaccagggc tacacacgtg ctacaatggc gtaaacaaag ggaggcgaac ccgcgagggt 1261 gggcaaatcc caaaaataac gtctcagttc ggattgtagt ctgcaactcg actacatgaa 1321 gctggaatcg ctagtaatcg cgaatcagaa tgtcgcggtg aatacgttcc cgggtcttgt 1381 acacaccgcc cgtcacacca tgggagtcag taacgcccga agccggtgac ccaacccgca 1441 agggagggag ccgtcgaagg tgggaccgat aactggggtg aagtcgtaac aaggtagccg 1501 tatcggaagg tgcggctgga tcacctcctt c

In certain embodiments, the intestinal microorganism comprises C. hiranonis, C. scindens or combination thereof. In certain embodiments, the intestinal microorganism comprises C. hiranonis. In certain embodiments, the intestinal microorganism comprises C. scindens.

By “percentage of identity” between two nucleic acid or amino acid sequences in the sense of the present disclosure, it is intended to indicate a percentage of nucleotides or of identical amino acid residues between the two sequences to be compared, obtained after the best alignment (optimum alignment), this percentage being purely statistical and the differences between the two sequences being distributed randomly and over their entire length. The comparisons of sequences between two nucleic acid or amino acid sequences are traditionally carried out by comparing these sequences after having aligned them in an optimum manner, said comparison being able to be carried out by segment or by “comparison window”. The optimum alignment of the sequences for the comparison can be carried out, in addition to manually, by means of the local homology algorithm of Smith and Waterman (1981) [Ad. App. Math. 2:482], by means of the local homology algorithm of Neddleman and Wunsch (1970) [J. Mol. Biol. 48: 443], by means of the similarity search method of Pearson and Lipman (1988) [Proc. Natl. Acad. Sci. USA 85:2444), by means of computer software using these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis., or else by BLAST N or BLAST P comparison software).

The percentage of identity between two nucleic acid or amino acid sequences is determined by comparing these two sequences aligned in an optimum manner and in which the nucleic acid or amino acid sequence to be compared can comprise additions or deletions with respect to the reference sequence for an optimum alignment between these two sequences. The percentage of identity is calculated by determining the number of identical positions for which the nucleotide or the amino acid residue is identical between the two sequences, by dividing this number of identical positions by the total number of positions in the comparison window and by multiplying the result obtained by 100 in order to obtain the percentage of identity between these two sequences.

For example, it is possible to use the BLAST program, “BLAST 2 sequences” (Tatusova et al., “Blast 2 sequences—a new tool for comparing protein and nucleotide sequences”, FEMS Microbiol Lett. 174:247-250) available on the site www.ncbi.nlm.nih.gov, the parameters used being those given by default (in particular for the parameters “open gap penalty”: 5, and “extension gap penalty”: 2; the matrix chosen being, for example, the matrix “BLOSUM 62” proposed by the program), the percentage of identity between the two sequences to be compared being calculated directly by the program. It is also possible to use other programs such as “ALIGN” or “Megalign” (DNASTAR) software.

By amino acid sequence having at least about 80%, e.g., at least about 85%, at least about 90%, at least about 95% and at least about 98% identity with a reference amino acid sequence, those having, with respect to the reference sequence, certain modifications, in particular a deletion, addition or substitution of at least one amino acid, a truncation or an elongation are preferred. In the case of a substitution of one or more consecutive or nonconsecutive amino acid(s), the substitutions are preferred in which the substituted amino acids are replaced by “equivalent” amino acids. The expression “equivalent amino acids” is aimed here at indicating any amino acid capable of being substituted with one of the amino acids of the base structure without, however, essentially modifying the biological activities of the corresponding antibodies and such as will be defined later, especially in the examples. These equivalent amino acids can be determined either by relying on their structural homology with the amino acids which they replace, or on results of comparative trials of biological activity between the different antibodies capable of being carried out.

By way of non-limiting example, Table 1 represents the possibilities of substitution capable of being carried out without resulting in a profound modification of the biological activity of the corresponding modified antibody, the reverse substitutions being naturally envisageable under the same conditions.

TABLE 1 Original residue Substitution(s) Ala (A) Val, Gly, Pro Arg (R) Lys, His Asn (N) Gln Asp (D) Glu Cys (C) Ser Gln (Q) Asn Glu (G) Asp Gly (G) Ala His (H) Arg Ile (I) Leu Leu (L) Ile, Val, Met Lys (K) Arg Met (M) Leu Phe (F) Tyr Pro (P) Ala Ser (S) Thr, Cys Thr (T) Ser Trp (W) Tyr Tyr (Y) Phe, Trp Val (V) Leu, Ala

Intestinal Microorganism Indicating Intestinal Health

In certain embodiments, the intestinal microorganism can be used to indicate intestinal health in a subject. In certain embodiments, the intestinal microorganism is associated with an intestinal disorder. In certain embodiments, the intestinal microorganism is associated with a heathy intestinal status. In certain embodiments, the intestinal microorganism more abundant in a healthy subject compared to a subject having an intestinal disorder. In certain embodiments, the intestinal microorganism less abundant in a healthy subject compared to a subject having an intestinal disorder.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more phylum selected from the group consisting of Actinobacteria, Bacteroidetes, Euryarchaeota, Firmicutes, Fusobacteria, Proteobacteria and Spirochaetae.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more class selected from the group consisting of Actinobacteria, Bacilli, Bacteroidia, Clostridia, Coriobacteria, Erysipelotrichia, Fusobacteria, Gammaproteobacteria, Methanobacteria, and Spirochaetes.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more order selected from the group consisting of Bacteriodales, Clostridiales, Coriobacteriales, Corynebacteriales, Enterobacteriales, Erysipelotrichales, Fusobacteriales, Lactobacillaes, Methanobacteriales and Spirochaetales.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more family selected from the group consisting of Bacteroidaceae, Clostridiaceae 1, Coriobacteriaceae, Corynebacteriaceae, Enterobacteriaceae, Erysipelotrichaceae, Fusobacteriaceae, Lachnospiraceae, Methanobacteriaceae, Peptostreptococcaceae, Porphyromonadaceae, Prevotellaceae, Ruminococcaceae, Spirochaetaceae, and Streptococcaceae.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more genus selected from the group consisting of Ruminococcus, Alloprevotella, Allisonella, Anaerostipes, Anaerobiospirillum, Bacteroides, Blautia, Clostridium sensu stricto 1, Collinsella, Coprococcus 1, Corynebacterium 1, Campylobacter, Enterococcus, Erysipelatoclostridium, Escherichia-Shigella, Faecalitalea, Fusobacterium, Helicobacter, Intestinibacter, Lachnoclostridium, Lactobacillus, Megasphaera, Methanobrevibacter, Parabacteroides, Porphyromonas, Phascolarctobacterium, Peptoclostridium, Prevotellaceae UCG-001, Pseudocitrobacter, Ruminiclostridium 9, Sarcina, Streptococcus, Succinivibrio, Treponema 2, Turicibacter, Tyzzerella, and Tyzzerella 4.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more species selected from the group consisting of Enterococcus durans, E. coli and C. perfringens. In certain embodiments, the intestinal microorganism comprises E. coli and C. perfringens.

In certain embodiments, the intestinal microorganism is selected from the group consisting of C. hiranonis, C. scindens, Veillonellaceae, Streptococcaceae, Bacteroides, Fusobacterium, Collinsella, Sarcina, Clostridium sensu stricto 1, Faecalitalea, Streptococcus, Erysipelatoclostridium, Megasphaera, Blautia, Alloprevotella, Peptoclostridium, and any combination thereof. In certain embodiments, the intestinal microorganism is C. hiranonis, C. scindens or combination thereof.

In certain embodiments, the intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to any sequence in Table 11.

In certain embodiments, the intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to the 16S rRNA nucleotide sequence of HQ802983.1.1440, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, New.ReferenceOTU82, GQ449092.1.1375, FJ506371.1.1371, GQ448744.1.1393, FJ957494.1.1454, HQ760911.1.1437, GQ006324.1.1342, GQ448246.1.1389, KC245406.1.1465, New.ReferenceOTU54, HQ751549.1.1448, JF712675.1.1540, JQ208181.1.1352, GX182404.8.1529, FP929060.3837.5503, FN667392.1.1495, FN667422.1.1495, HK557089.3.1395, HQ803964.1.1435, AM276759.1.1484, HK555938.1.1357, KF842598.1.1394, HQ792778.1.1436, FM865905.1.1392, FN563300.1.1447, HQ754680.1.1441, GQ867426.1.1494, EU470512.1.1400, AY239462.1.1500, New.ReferenceOTU114, FN668375.4306350.4307737, AB009242.1.1451, HQ792787.1.1438, AB506370.1.1516, DQ057365.1.1393, FN667084.1.1493, DQ113765.1.1450, HK694029.9.1487, AJ270486.1.1241, EU768569.1.1352, FM179752.1.1686, FJ957528.1.1445, KC504009.1.1465, GQ448506.1.1374, JF224013.1.1362, EU774020.1.1361, GQ448486.1.1387, HQ793763.1.1451, JN387556.1.1324, or New.ReferenceOTU109 in Table 11.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more Operational Taxonomic Units (OTUs) selected from the group consisting of HQ802983.1.1440, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, New.ReferenceOTU82, GQ449092.1.1375, FJ506371.1.1371, GQ448744.1.1393, FJ957494.1.1454, HQ760911.1.1437, GQ006324.1.1342, GQ448246.1.1389, KC245406.1.1465, New.ReferenceOTU54, HQ751549.1.1448, JF712675.1.1540, JQ208181.1.1352, GX182404.8.1529, FP929060.3837.5503, FN667392.1.1495, FN667422.1.1495, HK557089.3.1395, HQ803964.1.1435, AM276759.1.1484, HK555938.1.1357, KF842598.1.1394, HQ792778.1.1436, FM865905.1.1392, FN563300.1.1447, HQ754680.1.1441, GQ867426.1.1494, EU470512.1.1400, AY239462.1.1500, New.ReferenceOTU114, FN668375.4306350.4307737, AB009242.1.1451, HQ792787.1.1438, AB506370.1.1516, DQ057365.1.1393, FN667084.1.1493, DQ113765.1.1450, HK694029.9.1487, AJ270486.1.1241, EU768569.1.1352, FM179752.1.1686, JF807116.1.1260, FJ957528.1.1445, KC504009.1.1465, GQ448506.1.1374, JF224013.1.1362, EU774020.1.1361, GQ448486.1.1387, HQ793763.1.1451, JN387556.1.1324, and New.ReferenceOTU109.

In certain embodiments, the intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to the 16S rRNA nucleotide sequence of JRPJ01000002.1034290.1035971, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU45, HK555938.1.1357, FJ957494.1.1454, New.ReferenceOTU52, DQ797046.1.1403, GQ449092.1.1375, AMCI01001631.34.1456, KF842598.1.1394, HQ793763.1.1451, DQ113765.1.1450, ACBW01000012.3536.5054, HK693629.1.1491, JQ208053.1.1336, GQ493166.1.1359, GQ448486.1.1387, GQ491426.1.1332, New.ReferenceOTU54, or JN387556.1.1324 in Table 11.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more Operational Taxonomic Units (OTUs) selected from the group consisting of JRPJ01000002.1034290.1035971, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU45, HK555938.1.1357, FJ957494.1.1454, New.ReferenceOTU52, DQ797046.1.1403, GQ449092.1.1375, AMCI01001631.34.1456, KF842598.1.1394, HQ793763.1.1451, DQ113765.1.1450, ACBW01000012.3536.5054, HK693629.1.1491, JQ208053.1.1336, GQ493166.1.1359, GQ448486.1.1387, GQ491426.1.1332, New.ReferenceOTU54, and JN387556.1.1324.

In certain embodiments, the intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to the 16S rRNA nucleotide sequence of GQ006324.1.1342, New.ReferenceOTU52, HG798451.1.1400, HK557089.3.1395, GQ448336.1.1418, KF842598.1.1394, FJ950694.1.1472, HQ802983.1.1440, GQ448468.1.1366, or JN387556.1.1324 in Table 11.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more Operational Taxonomic Units (OTUs) selected from the group consisting of GQ006324.1.1342, New.ReferenceOTU52, HG798451.1.1400, HK557089.3.1395, GQ448336.1.1418, KF842598.1.1394, FJ950694.1.1472, HQ802983.1.1440, GQ448468.1.1366, and JN387556.1.1324.

In certain embodiments, the intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to the 16S rRNA nucleotide sequence of JRPJ01000002.1034290.1035971, New.ReferenceOTU45, GQ006324.1.1342, HK555938.1.1357, FJ957551.1.1489, FJ957494.1.1454, New.ReferenceOTU52, FM865905.1.1392, GQ016239.1.1362, HG798451.1.1400, EU461791.1.1414, GU303759.1.1517, New.ReferenceOTU114, AB506154.1.1541, EU774370.1.1398, HK557089.3.1395, HQ807346.1.1456, HQ748204.1.1442, GU179917.1.1382, GQ448336.1.1418, DQ804865.1.1390, GQ491757.1.1361, New.ReferenceOTU56, KF842598.1.1394, HQ802052.1.1445, GX182404.8.1529, FJ950694.1.1472, GQ448506.1.1374, HQ802983.1.1440, DQ793824.1.1370, GQ448468.1.1366, EU774020.1.1361, GQ491183.1.1360, GQ491426.1.1332, GQ493039.1.1311, JN387556.1.1324, and EU775983.1.1288 in Table 11.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more Operational Taxonomic Units (OTUs) selected from the group consisting of JRPJ01000002.1034290.1035971, New.ReferenceOTU45, GQ006324.1.1342, HK555938.1.1357, FJ957551.1.1489, FJ957494.1.1454, New.ReferenceOTU52, FM865905.1.1392, GQ016239.1.1362, HG798451.1.1400, EU461791.1.1414, GU303759.1.1517, New.ReferenceOTU114, AB506154.1.1541, EU774370.1.1398, HK557089.3.1395, HQ807346.1.1456, HQ748204.1.1442, GU179917.1.1382, GQ448336.1.1418, DQ804865.1.1390, GQ491757.1.1361, New.ReferenceOTU56, KF842598.1.1394, HQ802052.1.1445, GX182404.8.1529, FJ950694.1.1472, GQ448506.1.1374, HQ802983.1.1440, DQ793824.1.1370, GQ448468.1.1366, EU774020.1.1361, GQ491183.1.1360, GQ491426.1.1332, GQ493039.1.1311, JN387556.1.1324, and EU775983.1.1288.

In certain embodiments, the intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to the 16S rRNA nucleotide sequence of GQ449137.1.1391, HK555938.1.1357, GQ358246.1.1466, New.ReferenceOTU82, New.ReferenceOTU52, GQ138615.1.1402, JN681884.1.1409, GU303759.1.1517, New.ReferenceOTU114, EU774881.1.1422, AB469559.1.1551, HK557089.3.1395, EU358719.1.1513, HQ748204.1.1442, GQ338727.1.1397, HQ803964.1.1435, FJ951866.1.1493, EU772870.1.1289, GQ448468.1.1366, EU774020.1.1361, HQ782658.1.1415, DQ794633.1.1395, FN668375.4306350.4307737, or GQ867445.1.1457 in Table 11.

In certain embodiments, the intestinal microorganism comprises one or more bacteria and/or archaea of one or more Operational Taxonomic Units (OTUs) selected from the group consisting of GQ449137.1.1391, HK555938.1.1357, GQ358246.1.1466, New.ReferenceOTU82, New.ReferenceOTU52, GQ138615.1.1402, JN681884.1.1409, GU303759.1.1517, New.ReferenceOTU114, EU774881.1.1422, AB469559.1.1551, HK557089.3.1395, EU358719.1.1513, HQ748204.1.1442, GQ338727.1.1397, HQ803964.1.1435, FJ951866.1.1493, EU772870.1.1289, GQ448468.1.1366, EU774020.1.1361, HQ782658.1.1415, DQ794633.1.1395, FN668375.4306350.4307737, and GQ867445.1.1457.

Health Assessment Tools

The presently disclosed subject matter further provides a health assessment tool relating to the microorganisms disclosed herein. In certain embodiments, the health assessment tool is for monitoring intestinal health status or dysbiosis. In certain embodiments, the health assessment tool comprises one or more probe for detecting an amount of one or more microorganisms disclosed herein. In certain embodiments, the health assessment tool comprises a microarray of one or more probe for detecting an amount of one or more microorganism disclosed herein. In certain embodiments, the probe comprises a nucleic acid probe for detecting a signature gene of a microorganism disclosed herein. In certain embodiments, the probe detects a 16S rRNA sequence of a microorganism disclosed herein. In certain embodiments, the probe comprises an antibody. In certain embodiments, the antibody binds to a surface protein/antigen of a microorganism disclosed herein.

In certain embodiments, the amount of the microorganism is measured from a fecal sample of the subject. In certain embodiments, the health assessment tool monitoring intestinal health status or dysbiosis by comparing the amount of the one or more microorganism with a reference amount of the one or more microorganism.

In certain embodiments, the health assessment tool comprises probes for detecting at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 12, at least about 14, at least about 26 or more microorganisms disclosed herein. In certain embodiments, the health assessment tool comprises probes for detecting about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 12, about 14, or about 26 microorganisms disclosed herein. In certain embodiments, the health assessment tool comprises probes for detecting between about 1 to about 500, between about 1 to about 100, between about 1 to about 26, between about 5 to about 100, between about 5 to about 26, between about 10 to about 26, between about 15 to about 50, or between about 50 to about 100 microorganisms disclosed herein.

In certain embodiments, the one or more microorganism comprises a bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%) homologous or identical to any sequence in Table 11.

3. Pharmaceutical Composition

The presently disclosed subject matter provides a pharmaceutical composition for use as a medicament. In certain embodiments, the pharmaceutical composition comprises an effective amount of a bacterium capable of producing a first bile acid. In certain embodiments, the pharmaceutical composition further comprises an effective amount of a second bile acid. In certain embodiments, the bacterium is any bacterium disclosed in the above section. In certain embodiments, the first bile acid and/or the second bile acid is any bile acid disclosed in the above section or a pharmaceutically acceptable salt thereof. In certain embodiments, the first bile acid and the second bile acid are the same. In certain embodiments, the first bile acid and the second bile acid are different.

In certain embodiments, the bacterium comprised in the pharmaceutical composition is between about 1 thousand CFU and about 100 trillion CFU. In certain embodiments, the bacterium is between about 1 thousand CFU and about 1 trillion CFU, between about 1 million CFU and about 1 trillion CFU, between about 100 million CFU and about 100 billion CFU, between about 1 billion CFU and about 1 trillion CFU, between about 1 billion CFU and about 100 billion CFU, between about 100 million CFU and about 100 billion CFU, between about 1 billion CFU and about 50 billion CFU, between about 100 million CFU and about 50 billion CFU, or between about 1 billion CFU and about 10 billion CFU. In certain embodiments, the bacterium comprised in the pharmaceutical composition is at least about 1 thousand CFU, at least about 1 million CFU, at least about 10 million CFU, at least about 100 million CFU, at least about 1 billion CFU, at least about 10 billion CFU, at least about 100 billion CFU or more.

In certain embodiments, the second bile acid comprised in the pharmaceutical composition is between about 1 μg/unit dose and about 1 g/unit dose. In certain embodiments, the second bile acid comprised in the pharmaceutical composition is between about 10 μg/unit dose and about 1 g/unit dose, between about 10 μg/unit dose and about 500 mg/unit dose, between about 100 μg/unit dose and about 500 mg/unit dose, between about 1 mg/unit dose and about 500 mg/unit dose, between about 10 mg/unit dose and about 500 mg/unit dose, between about 100 mg/unit dose and about 500 mg/unit dose, between about 10 mg/unit dose and about 100 mg/unit dose, between about 50 mg/unit dose and about 300 mg/unit dose. In certain embodiments, the second bile acid comprised in the pharmaceutical composition is at least about 1 μg/unit dose, at least about 10 μg/unit dose, at least about 100 μg/unit dose, at least about 1 mg/unit dose, at least about 10 mg/unit dose, at least about 100 mg/unit dose, at least about 1 g/unit dose or more

The presently disclosed subject matter provides a bile acid for the treatment of an intestinal disorder in a dog. In certain embodiments, the bile acid is selected from the group consisting of chenodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, taurochenodeoxycholic acid, taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid and any combination thereof. In certain embodiments, the bile acid is a secondary bile acid. In certain embodiments, the secondary bile acid is selected from the group consisting of taurodeoxycholic acid, glycodeoxycholic acid, ursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholic acid, taurolithocholic acid, alpha-muricholic acid, deoxycholic acid, gamma-muricholic acid, glycolithocholic acid, taurolithocholic acid, lithocholic acid, omega-muricholic acid and any combination thereof. In certain embodiments, the secondary bile acid is deoxycholic acid and/or lithocholic acid.

In certain embodiments, the pharmaceutical composition is for the treatment of an intestinal disorder in a subject in need thereof. In certain embodiments, the intestinal disorder is selected from the ground consisting of irritable bowel syndrome, constipation, gastritis, colitis, inflammatory bowel disease (IBD), gastrointestinal ulcers, haemorrhagic gastroenteritis, diarrhea, Crohn's disease, ulcerative colitis, enteritis, antibiotic associated diarrhea, acute or chronic enteropathy, necrotizing enterocoloitis, and any combination thereof.

In certain embodiments, the subject is a dog. In certain embodiments, the intestinal disorder is an acute enteropathy or a chronic enteropathy. In certain embodiments, the intestinal disorder is a chronic enteropathy selected from the group consisting of food responsive enteropathy, antibiotic responsive enterophaty, and idiophathic inflammatory bowel disease (IBD).

In certain non-limiting embodiments, the subject is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a companion animal is a feline (e.g., a domestic cat) or a canine (e.g., a domestic dog).

The exact dose and frequency of administration depends on the particular condition being treated, the age, weight and general physical condition of the particular patient as well as other medication the individual can be taking, as is well known to those skilled in the art. Generally, the daily dose of a pharmaceutical composition disclosed herein can be in the range of between about 0.01 mg to about 1000 mg/day. In certain embodiments, the pharmaceutical composition can be about 0.05 mg to about 1000 mg/day, about 0.1 mg to about 1000 mg/day, about 1 mg to about 500 mg/day, about 0.01 mg to about 500 mg/day, about 0.05 mg to about 200 mg/day, about 1 mg to about 500 mg/day, about 1 mg to about 200 mg/day, about 5 mg to about 500 mg/day, about 50 mg to about 200 mg/day, about 100 mg to about 200 mg/day, about 100 mg to about 1000 mg/day, about 20 mg to about 50 mg/day, or about 20 mg to about 100 mg/day.

In certain embodiments, the pharmaceutical composition disclosed herein can be administered from about 10 times per day to about once per day, from about 5 times per day to about once per day, or from about thrice per day to about once per day. In certain embodiments, the pharmaceutical composition disclosed herein can be administered once per day. In certain embodiments, the pharmaceutical composition disclosed herein can be administered once per two days, once per three days, once per four days, once per five days, once per six days, once a week, once per two weeks, once per three weeks, or once per month.

The pharmaceutical composition disclosed herein can be administered in a variety of forms. In certain embodiments, the pharmaceutical composition disclosed herein can be administered orally, parenterally, rectally. In certain embodiments, orally administered pharmaceutical composition in solid dosage forms can be administered as capsules, dragees, granules, pills, powders, and tablets. In certain embodiments, the pharmaceutical composition can be administered in liquid form as elixirs, emulsions, microemulsions, solutions, suspensions, and syrups. In certain embodiments, parenterally administered pharmaceutical composition can be administered as aqueous or oleaginous solutions or aqueous or oleaginous suspensions, which suspensions comprise crystalline, amorphous, or otherwise insoluble forms of the pharmaceutical composition. In certain embodiments, rectally administered pharmaceutical composition can be administered as creams, gels, lotions, ointments, and pastes.

Depending upon the form of administration, the pharmaceutical composition disclosed herein can be formulated or administered with or without a pharmaceutically acceptable excipient. In certain embodiments, the excipients include encapsulating materials or formulation additives such as absorption accelerators, antioxidants, binders, buffers, coating agents, coloring agents, diluents, disintegrating agents, emulsifiers, extenders, fillers, flavoring agents, humectants, lubricants, perfumes, preservatives, propellants, releasing agents, sterilizing agents, sweeteners, solubilizers, wetting agents, solution aid, and any combination thereof. In certain embodiments, the pharmaceutical composition disclosed herein is administered without a solubilization aid. In certain embodiments, the pharmaceutical composition can separately be provided or packaged as kits.

4. Food Products

The presently disclosed subject matter provides a food product for improving intestinal health. In certain embodiments, the food product comprises an effective amount of a bacterium capable of producing a first bile acid. In certain embodiments, the food product further comprises an effective amount of a second bile acid. In certain embodiments, the bacterium is any bacterium disclosed in the above section. In certain embodiments, the first bile acid and/or the second bile acid is any bile acid disclosed in the above section or an edible salt thereof. In certain embodiments, the first bile acid and the second bile acid are the same. In certain embodiments, the first bile acid and the second bile acid are different.

In certain embodiments, the food product is a dietary supplement. In certain embodiments, the food product is a human food product. In certain embodiments, the food product is a pet food product, e.g., a cat food product or a dog food product. In certain embodiments, the food product is a dog food product. In certain embodiments, the food product is a pet dietary supplement.

In certain embodiments, the bacterium comprised in the pharmaceutical composition is between about 10 thousand CFU and about 100 trillion CFU. In certain embodiments, the bacterium is between about 1 thousand CFU and about 1 trillion CFU, between about 1 million CFU and about 1 trillion CFU, between about 100 million CFU and about 100 billion CFU, between about 1 billion CFU and about 1 trillion CFU, between about 1 billion CFU and about 100 billion CFU, between about 100 million CFU and about 100 billion CFU, between about 1 billion CFU and about 50 billion CFU, between about 100 million CFU and about 50 billion CFU, or between about 1 billion CFU and about 10 billion CFU. In certain embodiments, the bacterium comprised in the pharmaceutical composition is at least about 1 thousand CFU, at least about 1 million CFU, at least about 10 million CFU, at least about 100 million CFU, at least about 1 billion CFU, at least about 10 billion CFU, at least about 100 billion CFU or more.

In certain embodiments, the second bile acid comprised in the pharmaceutical composition is between about 1 μg/daily serving dose and about 1 g/daily serving dose. In certain embodiments, the second bile acid comprised in the pharmaceutical composition is between about 10 μg/daily serving dose and about 1 g/daily serving dose, between about 10 μg/daily serving dose and about 500 mg/daily serving dose, between about 100 μg/daily serving dose and about 500 mg/daily serving dose, between about 1 mg/daily serving dose and about 500 mg/daily serving dose, between about 10 mg/daily serving dose and about 500 mg/daily serving dose, between about 100 mg/daily serving dose and about 500 mg/daily serving dose, between about 10 mg/daily serving dose and about 100 mg/daily serving dose, between about 50 mg/daily serving dose and about 300 mg/daily serving dose. In certain embodiments, the second bile acid comprised in the pharmaceutical composition is at least about 1 μg/daily serving dose, at least about 10 μg/daily serving dose, at least about 100 μg/daily serving dose, at least about 1 mg/daily serving dose, at least about 10 mg/daily serving dose, at least about 100 mg/daily serving dose, at least about 1 g/daily serving dose or more.

In certain embodiments, a formulation of the presently disclosed subject matter can further comprise an additional active agent. Non-limiting examples of additional active agents that can be present within a formulation of the presently disclosed subject matter include a nutritional agent (e.g., amino acids, peptides, proteins, fatty acids, carbohydrates, sugars, nucleic acids, nucleotides, vitamins, minerals, etc.), a prebiotic, a probiotic, an antioxidant, and/or an agent that improves animal health.

In certain embodiments, the food product comprises one or more probiotic. In certain embodiments, the probiotic is a human probiotic. In certain embodiments, the probiotic is an animal probiotic. In certain embodiments, the animal probiotic is a feline probiotic. In certain embodiments, the animal probiotic is a canine probiotic. In certain embodiments, the probiotic is bifidobacterium, lactic acid bacterium and/or enterococcus. In certain embodiments, the probiotic is selected from the group consisting of any organism from lactic acid bacteria and more specifically from the following bacterial genera; Lactococcus spp., Pediococcus spp., Bifidobacterium spp. (e.g., B. longum B. bifidum, B. pseudolongum, B. animalis), Lactobacillus spp. (e.g. L. bulgaricus, L. acidophilus, L. brevis, L casei, L. rhamnosus, L. plantarum, L. reuteri, L. fermentum, Enterococcus spp. (e.g. E. faecium), Prevotella spp., Fusobacteria spp, Alloprevotella spp, and any combination thereof. In certain embodiments, the probiotic is administered to a companion animal in an amount of from about 1 colony forming unit (CFU) to about 100 billion CFUs per day for the maintenance of GI microflora. In certain embodiments, the probiotic is administered to a companion animal in an amount of from about 1 colony forming unit (CFU) to about 20 billion CFUs per day for the maintenance of GI microflora. In certain embodiments, the probiotic is administered to a companion animal in an amount of from about 1 billion CFUs to about 20 billion CFUs per day for the maintenance of GI microflora. In certain embodiments, the probiotic is administered to a companion animal in amounts of from about 0.01 billion to about 100 billion live bacteria per day. In certain embodiments, the probiotic is administered to a companion animal in amounts of from about 0.1 billion to about 10 billion live bacteria per day.

In certain embodiments, an additional prebiotic can be included, such as fructooligosaccharides (FOS), xylooligosaccharides (XOS), galactooligosaccharides (GOS), glucans, galactans, arabinogalactan, inulin and/or mannooligosaccharides. In certain embodiments, the additional prebiotic is administered in amounts sufficient to positively stimulate the GI microflora and/or cause one or more probiotics to proliferate.

In certain embodiments, the companion animal food product can further contain additives known in the art. In certain embodiments, such additives are present in amounts that do not impair the purpose and effect provided by the presently disclosed subject matter. Examples of contemplated additives include, but are not limited to, substances that are functionally beneficial to improving health, substances with a stabilizing effect, organoleptic substances, processing aids, substances that enhance palatability, coloring substances, and substances that provide nutritional benefits. In certain embodiments, the stabilizing substances include, but are not limited to, substances that tend to increase the shelf life of the product. In certain embodiments, such substances include, but are not limited to, preservatives, synergists and sequestrants, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and humectants. In certain embodiments, the emulsifiers and/or thickening agents include, for example, gelatin, cellulose ethers, starch, starch esters, starch ethers, and modified starches.

In certain embodiments, the additives for coloring, palatability, and nutritional purposes include, for example, colorants; iron oxide, sodium chloride, potassium citrate, potassium chloride, and other edible salts; vitamins; minerals; and flavoring. The amount of such additives in a product typically is up to about 5% (dry basis of the product).

In certain embodiments, the companion animal food product is a dietary supplement. In certain embodiments, the dietary supplements include, for example, a feed used with another feed to improve the nutritive balance or performance of the total. In certain embodiments, the supplements include compositions that are fed undiluted as a supplement to other feeds, offered free choice with other parts of an animal's ration that are separately available, or diluted and mixed with an animal's regular feed to produce a complete feed. The AAFCO, for example, provides a discussion relating to supplements in the American Feed Control Officials, Incorp. Official Publication, p. 220 (2003). Supplements can be in various forms including, for example, powders, liquids, syrups, pills, tablets, encapsulated compositions, etc.

In certain embodiments, the companion animal food product is a treat. In certain embodiments, treats include, for example, compositions that are given to an animal to entice the animal to eat during a non-meal time. In certain embodiments, the companion animal food product is a treat for canines include, for example, dog bones. Treats can be nutritional, wherein the product comprises one or more nutrients, and can, for example, have a composition as described above for food. Non-nutritional treats encompass any other treats that are non-toxic.

In certain embodiments, a bacterium and/or a bile acid of the presently disclosed subject matter can be incorporated into the composition during the processing of the formulation, such as during and/or after mixing of other components of the product. Distribution of these components into the product can be accomplished by conventional means.

In certain embodiments, companion animal food products of the presently disclosed subject matter can be prepared in a canned or wet form using conventional companion animal food processes. In certain embodiments, ground animal (e.g., mammal, poultry, and/or fish) proteinaceous tissues are mixed with the other ingredients, such as milk fish oils, cereal grains, other nutritionally balancing ingredients, special purpose additives (e.g., vitamin and mineral mixtures, inorganic salts, cellulose and beet pulp, bulking agents, and the like); and water that sufficient for processing is also added. These ingredients are mixed in a vessel suitable for heating while blending the components. Heating of the mixture can be effected using any suitable manner, such as, for example, by direct steam injection or by using a vessel fitted with a heat exchanger. Following the addition of the last ingredient, the mixture is heated to a temperature range of from about 50° F. to about 212° F. Temperatures outside this range are acceptable but can be commercially impractical without use of other processing aids. When heated to the appropriate temperature, the material will typically be in the form of a thick liquid. The thick liquid is filled into cans. A lid is applied, and the container is hermetically sealed. The sealed can is then placed into conventional equipment designed to sterilize the contents. This is usually accomplished by heating to temperatures of greater than about 230° F. for an appropriate time, which is dependent on, for example, the temperature used and the composition.

In certain embodiments, companion animal food products of the presently disclosed subject matter can be prepared in a dry form using conventional processes. In certain embodiments, dry ingredients, including, for example, animal protein sources, plant protein sources, grains, etc., are ground and mixed together. In certain embodiments, moist or liquid ingredients, including fats, oils, animal protein sources, water, etc., are then added to and mixed with the dry mix. In certain embodiments, the mixture is then processed into kibbles or similar dry pieces. In certain embodiments, the companion animal food product is kibble. In certain embodiments, kibble is formed using an extrusion process in which the mixture of dry and wet ingredients is subjected to mechanical work at a high pressure and temperature and forced through small openings and cut off into kibble by a rotating knife. In certain embodiments, the wet kibble is then dried and optionally coated with one or more topical coatings which can include, for example, flavors, fats, oils, powders, and the like. In certain embodiments, kibble can also be made from the dough using a baking process, rather than extrusion, wherein the dough is placed into a mold before dry-heat processing.

In certain embodiments, treats of the presently disclosed subject matter can be prepared by, for example, an extrusion or baking process similar to those described above for dry food.

The presently disclosed subject matter provides a diet for increase a population of a bacterium capable of producing a bile acid in a companion animal. In certain embodiments, the diet comprises protein, fat, crude fiber, total dietary fiber, carbohydrate, calcium, phosphorus, sodium, chloride, potassium, magnesium, iron, copper, manganese, zinc, iodine, selenium, vitamin A, vitamin D3, vitamin E, vitamin C, thiamine (vitamin B1), riboflavin (vitamin B2), pantothenic acid, niacin, pyridoxine (vitamin B6), folic acid, biotin, cobalannin (vitamin B12), choline, arginine, lysine, methionine, cystine, taurine, linoleic acid, arachidonic acid, Omega-6 fatty acids, Omega-3 fatty acids, EPA, and/or DHA.

In certain embodiments, the subject is a dog. In certain embodiments, the diet is a Royal Canin Veterinary Diet. In certain embodiments, the diet is selected from the group consisting of Ultamino, Hydrolyzed Protein Adult HP Dry, Hydrolyzed Protein Wet, Hydrolyzed Protein Adult PS Dry, Hydrolyzed Protein Moderate Calorie Dry, Hydrolyzed Protein Small Dog Dry, Hydrolyzed protein Treats, and any combination thereof.

In certain embodiments, the bacterium comprises a bile acid-inducible operon (bai operon). In certain embodiments, the bacterium is C. hiranonis, C. scindens or combination thereof. In certain embodiments, the bacterium is C. hiranonis.

The presently disclosed subject matter provides a Royal Canin Veterinary Diet for the treatment of an intestinal disorder in a dog, wherein the dog comprises a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism, and wherein the first amount of the first intestinal microorganism is higher than a first reference amount of the first intestinal microorganism, and/or the second amount of the second intestinal microorganism is lower than a second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of New.ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, JQ208053.1.1336, and any combination thereof. In certain embodiments, the second intestinal microorganism is selected from the group consisting of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, HK555938.1.1357, and any combination thereof.

5. Treatment Methods

In certain non-limiting embodiments, the presently disclosed subject matter provides for a method for improving intestinal health and/or treating an intestinal disorder of a subject in need thereof. In certain embodiments, the method can improve immunity, digestive function and/or decrease inflammation of a companion animal.

In certain non-limiting embodiments, the presently disclosed subject matter provides for a method for determining susceptibility of an intestinal disorder in a companion animal. In certain embodiments, the method comprises:

a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal;

b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and

c) determining that the companion animal is susceptible of an intestinal disorder, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HQ802983.1.1440, GQ449092.1.1375, GQ448744.1.1393, KF842598.1.1394, HG798451.1.1400, New.ReferenceOTU52, HK555938.1.1357, FJ957494.1.1454, FN667392.1.1495, New.ReferenceOTU54, HQ760911.1.1437, GQ006324.1.1342, FJ950694.1.1472, FM865905.1.1392, FJ506371.1.1371, FJ957528.1.1445, JF712675.1.1540, New.ReferenceOTU82, AB009242.1.1451, HQ751549.1.1448, AB506370.1.1516, DQ057365.1.1393, FN667422.1.1495, AJ270486.1.1241, FN668375.4306350.4307737, GQ867426.1.1494, GX182404.8.1529, JF224013.1.1362, GQ448246.1.1389, JF807116.1.1260, KC245406.1.1465, FN667084.1.1493, EU470512.1.1400, EU768569.1.1352, AY239462.1.1500, KC504009.1.1465, FM179752.1.1686, New.ReferenceOTU114, HK557089.3.1395, JQ208181.1.1352, HQ803964.1.1435, AM276759.1.1484, JN387556.1.1324, GQ448486.1.1387, HK694029.9.1487, HQ754680.1.1441, FN563300.1.1447, FP929060.3837.5503, GQ448506.1.1374, Enterococcus durans, C. perfringens, or E. coli.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of HQ802983.1.1440, GQ449092.1.1375, GQ448744.1.1393, KF842598.1.1394, HG798451.1.1400, New.ReferenceOTU52, HK555938.1.1357, FJ957494.1.1454, FN667392.1.1495, New.ReferenceOTU54, HQ760911.1.1437, GQ006324.1.1342, FJ950694.1.1472, FM865905.1.1392, FJ506371.1.1371, FJ957528.1.1445, JF712675.1.1540, New.ReferenceOTU82, AB009242.1.1451, HQ751549.1.1448, AB506370.1.1516, DQ057365.1.1393, FN667422.1.1495, AJ270486.1.1241, FN668375.4306350.4307737, GQ867426.1.1494, GX182404.8.1529, JF224013.1.1362, GQ448246.1.1389, JF807116.1.1260, KC245406.1.1465, FN667084.1.1493, EU470512.1.1400, EU768569.1.1352, AY239462.1.1500, KC504009.1.1465, FM179752.1.1686, New.ReferenceOTU114, HK557089.3.1395, JQ208181.1.1352, HQ803964.1.1435, AM276759.1.1484, JN387556.1.1324, GQ448486.1.1387, HK694029.9.1487, HQ754680.1.1441, FN563300.1.1447, FP929060.3837.5503, GQ448506.1.1374, Enterococcus durans, C. perfringens, E. coli and any combination thereof.

In certain embodiments, the first intestinal microorganism is C. perfringens, E. coli and any combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of EU774020.1.1361, HQ793763.1.1451, HQ792787.1.1438, New.ReferenceOTU109, HQ792778.1.1436, or DQ113765.1.1450.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of EU774020.1.1361, HQ793763.1.1451, HQ792787.1.1438, New.ReferenceOTU109, HQ792778.1.1436, DQ113765.1.1450, and any combination thereof.

In certain embodiments, the method further comprises providing a customized recommendation of a treatment regimen, and/or further monitoring the intestinal microorganism, when the first amount of the first intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.

In certain non-limiting embodiments, the presently disclosed subject matter provides for a method for determining responsiveness of a companion animal having an intestinal disorder to a diet. In certain embodiments, the method comprises:

a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal;

b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and

c) determining that the companion animal is responsive to the diet, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism, or determining that the companion animal is non-responsive to the diet, when the first amount of the intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, JQ208053.1.1336, and any combination thereof. In certain embodiments, the second intestinal microorganism is selected from the group consisting of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, or HK555938.1.1357.

In certain embodiments, the first intestinal microorganism is selected from the group consisting of New.ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, JQ208053.1.1336, and any combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, or HK555938.1.1357.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, HK555938.1.1357, and any combination thereof.

In certain embodiments, the method further comprises administering the diet to the companion animal when companion animal is determined as responsive to the diet. In certain embodiments, the method further comprises administering the diet, a steroid and optionally an antibiotic to the companion animal when companion animal is determined as non-responsive to the diet.

In certain embodiments, the determination in step c) occurs before administering the diet or the diet, the steroid and optionally the antibiotic to the companion animal.

In certain non-limiting embodiments, the presently disclosed subject matter provides for a method for determining effectiveness of a diet for treating an intestinal disorder in a companion animal. In certain embodiments, the method comprises:

a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal before or after administering a diet to a companion animal for treating an intestinal disorder;

b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and

c) determining that the diet is effective for treating an intestinal disorder, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism, or determining that the diet is ineffective for treating an intestinal disorder, when the first amount of the intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.

In certain embodiments, the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK557089.3.1395, or GQ448336.1.1418. In certain embodiments, the first intestinal microorganism is selected from the group consisting of HK557089.3.1395, GQ448336.1.1418, and combination thereof.

In certain embodiments, the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of KF842598.1.1394, GQ006324.1.1342, HQ802983.1.1440, JN387556.1.1324, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, or GQ448468.1.1366.

In certain embodiments, the second intestinal microorganism is selected from the group consisting of KF842598.1.1394, GQ006324.1.1342, HQ802983.1.1440, JN387556.1.1324, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, GQ448468.1.1366, and any combination thereof.

In certain embodiments, the method further comprises administering the diet to the companion animal when companion animal is determined as responsive to the diet. In certain embodiments, the method further comprises administering the diet, a steroid and optionally an antibiotic to the companion animal when companion animal is determined as non-responsive to the diet.

In certain embodiments, the determination in step c) occurs before administering the diet or the diet, the steroid and optionally the antibiotic to the companion animal.

In certain embodiments, the reference amount of an intestinal microorganism derived from a mean amount of the intestinal microorganism in a plurality of healthy companion animals. In certain embodiments, the amount of the intestinal bacterium is measured from a fecal sample of the subject.

In certain embodiments, the method comprises administering to the subject an effective amount of a presently disclosed pharmaceutical composition, an effective amount of a presently disclosed food product, or any combination thereof. In certain embodiments, the method further comprises monitoring an intestinal microorganism in the subject. In certain embodiments, the intestinal microorganism is sampled from a fecal sample of the subject.

In certain embodiments, the intestinal microorganism is selected from the group consisting of Ruminococcus, Alloprevotella, Allisonella, Anaerostipes, Anaerobiospirillum, Bacteroides, Blautia, Clostridium sensu stricto 1, Collinsella, Coprococcus 1, Corynebacterium 1, Campylobacter, Enterococcus, Erysipelatoclostridium, Escherichia-Shigella, Faecalitalea, Fusobacterium, Helicobacter, Intestinibacter, Lachnoclostridium, Lactobacillus, Megasphaera, Methanobrevibacter, Parabacteroides, Porphyromonas, Phascolarctobacterium, Peptoclostridium, Prevotellaceae UCG-001, Pseudocitrobacter, Ruminiclostridium 9, Sarcina, Streptococcus, Succinivibrio, Treponema 2, Turicibacter, Tyzzerella, Tyzzerella 4 and any combination thereof.

In certain embodiments, the intestinal microorganism is selected from the group consisting of Escherichia-Shigella, Clostridium sensu stricto 1, Enterococcus, Fusobacterium and any combination thereof. In certain embodiments, the intestinal microorganism is E. coli, C. perfringens or combination thereof.

In certain embodiments, an amount of the intestinal bacterium is decreased after administration of the pharmaceutical composition. In certain embodiments, an amount of the intestinal bacterium is decreased within about 14 days after administration of the pharmaceutical composition. In certain embodiments, an amount of the intestinal bacterium is decreased within about 21 days, within about 14 days, within about 12 days, within about 10 days, within about 7 days, within about 6 days, within about 5 days, within about 4 days, within about 3 days, within about 2 days, or within about 1 day after administration of the pharmaceutical composition. In certain embodiments, an amount of the intestinal bacterium is decreased within about 1 day to about 21 days, within about 1 days to about 14 days, within about 3 days to about 14 days, within about 5 days to about 14 days, within about 7 days to about 14 days, within about 10 days to about 14 days, or within about 7 days to about 21 days after administration of the pharmaceutical composition.

In certain embodiments, the intestinal microorganism is selected from the group consisting of C. hiranonis, C. scindens, Veillonellaceae, Streptococcaceae, Bacteroides, Fusobacterium, Collinsella, Sarcina, Clostridium sensu stricto 1, Faecalitalea, Streptococcus, Erysipelatoclostridium, Megasphaera, Blautia, Alloprevotella, Peptoclostridium, and any combination thereof. In certain embodiments, the intestinal microorganism is C. hiranonis, C. scindens or combination thereof.

In certain embodiments, an amount of the intestinal microorganism is increased after administration of the pharmaceutical composition and/or the food product. In certain embodiments, the amount of the intestinal microorganism is increased within about 14 days after administration of the pharmaceutical composition and/or the food product. In certain embodiments, an amount of the intestinal bacterium is increased within about 21 days, within about 14 days, within about 12 days, within about 10 days, within about 7 days, within about 6 days, within about 5 days, within about 4 days, within about 3 days, within about 2 days, or within about 1 day after administration of the pharmaceutical composition. In certain embodiments, an amount of the intestinal bacterium is increased within about 1 day to about 21 days, within about 1 days to about 14 days, within about 3 days to about 14 days, within about 5 days to about 14 days, within about 7 days to about 14 days, within about 10 days to about 14 days, or within about 7 days to about 21 days after administration of the pharmaceutical composition.

In certain embodiments, the method comprises:

a) measuring a first amount of one or more intestinal microorganism in the subject;

b) administering a treatment regimen to the subject for treating the intestinal disorder;

c) measuring a second amount of the intestinal microorganism in the subject after step b); and

d) continuing administering the treatment regimen, when the second amount of the intestinal microorganism is reduced compared to the first amount of the intestinal microorganism.

In certain embodiments, the second amount of the intestinal microorganism is measured between about 7 days and about 14 days after step b). In certain embodiments, an amount of the intestinal microorganism is decreased within about 21 days, within about 14 days, within about 12 days, within about 10 days, within about 7 days, within about 6 days, within about 5 days, within about 4 days, within about 3 days, within about 2 days, or within about 1 day after step b). In certain embodiments, an amount of the intestinal bacterium is decreased within about 1 day to about 21 days, within about 1 days to about 14 days, within about 3 days to about 14 days, within about 5 days to about 14 days, within about 7 days to about 14 days, within about 10 days to about 14 days, or within about 7 days to about 21 days after step b).

In certain embodiments, the intestinal microorganism is measured from a fecal sample of the subject.

In certain embodiments, the method comprises:

a) measuring the first amount of one or more intestinal microorganism in the subject;

b) comparing the first amount of the intestinal microorganism with a reference amount of the intestinal microorganism, wherein the reference amount of the intestinal microorganism is determined based on the amount of the intestinal microorganism in a plurality of healthy subjects;

c) providing a customized recommendation of a treatment regimen, and/or further monitoring the intestinal microorganism, when the first amount of the intestinal microorganism is above the reference amount of the intestinal microorganism.

In certain embodiments, the method further comprises measuring a second amount of the intestinal microorganism in the subject after step c), and continuing the treatment regimen when the second amount of the intestinal microorganism is decreased compared to the first amount of the intestinal microorganism and is above the reference amount of the intestinal microorganism.

In certain embodiments, the second amount of the intestinal bacterium is measured between about 7 days and about 14 days after step c). In certain embodiments, an amount of the intestinal microorganism is decreased within about 21 days, within about 14 days, within about 12 days, within about 10 days, within about 7 days, within about 6 days, within about 5 days, within about 4 days, within about 3 days, within about 2 days, or within about 1 day after step b). In certain embodiments, an amount of the intestinal microorganism is decreased within about 1 day to about 21 days, within about 1 days to about 14 days, within about 3 days to about 14 days, within about 5 days to about 14 days, within about 7 days to about 14 days, within about 10 days to about 14 days, or within about 7 days to about 21 days after step c).

In certain embodiments, the intestinal microorganism is measured from a fecal sample of the subject.

In certain embodiments, the intestinal microorganism is selected from the group consisting of Ruminococcus, Alloprevotella, Allisonella, Anaerostipes, Anaerobiospirillum, Bacteroides, Blautia, Clostridium sensu stricto 1, Collinsella, Coprococcus 1, Corynebacterium 1, Campylobacter, Enterococcus, Erysipelatoclostridium, Escherichia-Shigella, Faecalitalea, Fusobacterium, Helicobacter, Intestinibacter, Lachnoclostridium, Lactobacillus, Megasphaera, Methanobrevibacter, Parabacteroides, Porphyromonas, Phascolarctobacterium, Peptoclostridium, Prevotellaceae UCG-001, Pseudocitrobacter, Ruminiclostridium 9, Sarcina, Streptococcus, Succinivibrio, Treponema 2, Turicibacter, Tyzzerella, Tyzzerella 4 and any combination thereof. In certain embodiments, the intestinal microorganism is selected from the group consisting of Escherichia-Shigella, Clostridium sensu stricto 1, Enterococcus, Fusobacterium and any combination thereof. In certain embodiments, the intestinal microorganism is E. coli, C. perfringens or combination thereof.

In certain embodiments, the treatment regimen comprises administering an effective amount of a presently disclosed pharmaceutical composition, an effective amount of a presently disclosed food product, or any combination thereof.

In certain non-limiting embodiments, the subject is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a companion animal is a feline (e.g., a domestic cat) or a canine (e.g., a domestic dog). In certain non-limiting embodiments, the companion animal is at risk of an intestinal disorder and/or inflammation. In certain non-limiting embodiments, the companion animal is not known to be at risk of an intestinal disorder and/or inflammation. In certain non-limiting embodiments, the companion animal has an intestinal disorder and/or inflammation. In certain non-limiting embodiments, the companion animal is not known to have an intestinal disorder and/or inflammation. In certain non-limiting embodiments, the companion animal is under a treatment for a digestive disorder and/or inflammation. In certain non-limiting embodiments, the treatment is a dietary therapy. In certain embodiments, the companion animal is a dog. In certain embodiments, the intestinal disorder is an acute enteropathy or a chronic enteropathy. In certain embodiments, the intestinal disorder is a chronic enteropathy selected from the group consisting of food responsive enteropathy, antibiotic responsive enterophaty, and idiophathic inflammatory bowel disease (IBD).

In certain embodiments, the pharmaceutical composition and/or the food product can be administered to a subject from 20 times per day to once per day, from 10 times per day to once per day, or from 5 times per day to once per day. In certain embodiments, the pharmaceutical composition and/or the food product can be administered to a subject once per day, twice per day, thrice per day, 4 times per day, 5 times per day, 6 times per day, 7 times per day, 8 times per day, 9 times per day, 10 or more times per day. In certain embodiments, the pharmaceutical composition and/or the food product can be administered to a subject once per two days, once per three days, once per four days, once per five days, once per six days, once a week, once per two weeks, once per three weeks, or once per month. In certain embodiments, the food product can be administered to an animal in a constant manner, e.g., where the animal grazes on a constantly available supply of the subject food product.

In certain embodiments, the dosage of the pharmaceutical composition is between about 1 mg/kg body weight per day and about 5000 mg/kg body weight per day. In certain embodiments, the dosage of the pharmaceutical composition is between about 5 mg/kg body weight per day and about 1000 mg/kg body weight per day, between about 10 mg/kg body weight per day and about 500 mg/kg body weight per day, between about 10 mg/kg body weight per day and about 250 mg/kg body weight per day, between about 10 mg/kg body weight per day and about 200 mg/kg body weight per day, between about 20 mg/kg body weight per day and about 100 mg/kg body weight per day, between about 20 mg/kg body weight per day and about 50 mg/kg body weight per day or any intermediate range thereof. In certain embodiments, the dosage of the pharmaceutical composition is at least about 1 mg/kg body weight per day, at least about 5 mg/kg body weight per day, at least about 10 mg/kg body weight per day, at least about 20 mg/kg body weight per day, at least about 50 mg/kg body weight per day, at least about 100 mg/kg body weight per day, at least about 200 mg/kg body weight per day or more. In certain embodiments, the dosage of the pharmaceutical composition is no more than about 5 mg/kg body weight per day, no more than about 10 mg/kg body weight per day, no more than about 20 mg/kg body weight per day, no more than about 50 mg/kg body weight per day, no more than about 100 mg/kg body weight per day, no more than about 200 mg/kg body weight per day, no more than about 500 mg/kg body weight per day or more.

In certain embodiments, the amount of the pharmaceutical composition and/or the food product decreases over the course of feeding a companion animal. In certain embodiments, the concentration of the pharmaceutical composition and/or the food product increases over the course of feeding a companion animal. In certain embodiments, the concentration of the pharmaceutical composition and/or the food product is modified based on the age of the companion animal.

6. Kits

The presently disclosed subject matter provides kits for treating and/or preventing an intestinal disorder in a subject. In certain embodiments, the kit comprises an effective amount of the presently disclosed pharmaceutical composition, dietary supplement, functional food, food product, diet or any combination thereof. In certain embodiments, the kit comprises a sterile container; such containers can be boxes, ampules, bottles, vials, tubes, bags, pouches, blister-packs, or other suitable container forms known in the art. Such containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding medicaments.

If desired, the pharmaceutical composition, dietary supplement, functional food, food product, and/or diet are provided together with instructions for administering the same to a subject having or at risk of developing an intestinal disorder. The instructions generally include information about the use of the pharmaceutical composition, dietary supplement, functional food, food product, diet for the treatment and/or prevention of an intestinal disorder. In certain embodiments, the instructions include at least one of the following: description of the therapeutic agent; dosage schedule and administration for treatment or prevention of an intestinal disorder or symptoms thereof; precautions; warnings; indications; counter-indications; over-dosage information; adverse reactions; animal pharmacology; clinical studies; and/or references. The instructions can be printed directly on the container (when present), or as a label applied to the container, or as a separate sheet, pamphlet, card, or folder supplied in or with the container.

Advantageously, the kit can be packaged in per use groupings such that, for example, a daily prescription of each component can be identified in order to enhance patient compliance. Sets of the pharmaceutical composition can be identified in a variety of ways. For example, in certain embodiments, a set of the pharmaceutical composition, dietary supplement, functional food, food product, diet can be identified on the package containing the same. In certain embodiments, external instructions can be provided with a set or sets of the pharmaceutical composition, dietary supplement, functional food, food product, diet that, for example, identify a grouping and instruct a patient/animal owner appropriate times to take the pharmaceutical composition, dietary supplement, functional food, food product, diet of the kit.

EXAMPLES

The presently disclosed subject matter will be better understood by reference to the following Example, which is provided as exemplary of the present disclosure, and not by way of limitation.

Example 1 Introduction

Although a wide range of environmental factors have been shown to influence the microbiome, diet is regarded as one of the most potent modulators of the composition and function of the gut-resident microbial community in healthy humans and other mammals^(7,8) and can act as both a risk factor and a treatment modality for IBD^(9,10). Epidemiologic data and studies in mice have shown that diets high in fat and/or low in fiber, as well as dietary additives such as emulsifiers, are either risk factors for IBD, or in some cases can directly compromise intestinal barrier function leading to disease¹¹⁻¹³. Diet can be also leveraged to treat IBD, with perhaps the clearest example of this being the use of exclusive enteral nutrition (EEN) as first-line therapy for pediatric Crohn's disease¹⁴. High remission rates (≥60%) are observed following EEN and, compared to corticosteroids, EEN achieves better patient growth along with a reduction in biomarkers of disease, such as fecal calprotectin and C-reactive protein¹⁵⁻¹⁸ Interestingly, EEN has a marked effect on the microbiome, but the precise nature of this effect has been complicated to discern, with some studies reporting reduced microbiome diversity following EEN therapy¹⁹⁻²¹ while others point to relatively unchanged^(22,23) or increased diversity²⁴.

The mechanisms by which diet impacts the gut microbiome to ameliorate IBD symptoms are unclear and are complicated to dissect from human subject research were diet is challenging to control, necessitating either retrospective studies in conjunction with extensive food intake surveys²⁵, controlled feeding studies²⁶, or focusing on populations with different subsistence practices²⁷⁻²⁹. In contrast, mouse models of colitis have yielded important insights into the pathophysiology of intestinal inflammation, but these often involve chemical or genetic perturbation, rather than spontaneous disease development. Moreover, the ubiquitous use of autoclaved food and acidified water for mouse husbandry, together with the tendency for cage effects to dominate in mouse microbiome studies, raises concerns about clinical relevance of diet-microbiome studies in these models of colitis. As companion animals, dogs share the same environment as humans, and spontaneously develop a chronic enteritis that clinically resembles human IBD, including similar gastrointestinal pathology, responsiveness to similar treatments^(30,31), involvement of some of the same susceptibility loci³⁰⁻³², and shared disease-associated microbial taxa³³⁻³⁵. Intriguingly, after treatment with dietary therapy, over 50% of dogs with chronic enteritis enter a long-lasting state of remission³⁶, making the use of prescription diets the first-line treatment for IBD in companion animal medicine. A recent metagenomic study produced a catalog of over one million taxonomically and functionally annotated microbial genes from the canine gut and showed that compared to other mammals, such as the mouse and pig—the microbial environment in dogs most closely resembles that of humans³⁷. Furthermore, the canine microbiome was markedly altered by diet change in a manner that resembles what has been reported in humans³⁷. Together, these data argue that dogs are an ideal animal model in which to study diet-microbiome interactions in the context of intestinal disease.

Despite the fact that the gut microbiome has been implicated in IBD pathogenesis, and that diet profoundly alters the microbiome and can be used to manage symptoms of IBD, there is limited insight into the mechanisms by which this occurs. In this study, treatment-naive dogs were examined with chronic enteritis and changes in their fecal microbial community structure and metabolites in response to treatment were monitored. By comparing changes over time in diet-responsive dogs, versus animals that failed diet therapy and required subsequent combination therapy, it was shown that diet induces rapid remission by shaping the community structure and re-programming the metabolic function of the microbiome. Notably, it was demonstrated that secondary bile acids, likely produced by clostridia, are involved in the diet induced alterations of microbiota community by inhibiting the growth of potential pathogens. These findings provide a general mechanism by which diet can modulate microbial communities to reduce GI disease.

Methods Diagnosis and Treatment of Canine Chronic Enteritis (CCE)

Client-owned animals presenting with clinical signs of CCE were screened at the Ryan Veterinary Hospital of the University of Pennsylvania. All animal work was carried out in accordance within the guidelines of the University of Pennsylvania IACUC (Protocol 805283), and signed owner consent was obtained before enrollment. Dogs were screened if they had any one of the following clinical signs for ≥3 weeks' duration: vomiting, diarrhea or weight loss despite adequate caloric intake. Dogs were excluded from screening if they had been treated with a hydrolyzed protein diet, antibiotics, corticosteroids or probiotics within the previous two weeks. At the time of screening, the following were performed on each animal: complete physical examination, routine fecal screening (including zinc sulfate flotation for parasite identification, gram stain and culture for Salmonella spp. and Campylobacter spp.), complete blood count, serum biochemical profile, serum measurement of canine trypsin-like immunoreactivity, cobalamin and folate, urinalysis, abdominal ultrasound examination, and disease severity scoring using the Canine Chronic Enteropathy Clinical Activity Index (CCECAI)³⁶. If these initial screening tests failed to identify a cause for the clinical signs, upper and/or lower gastrointestinal endoscopy with mucosal biopsies was performed. Biopsies were fixed in formalin, embedded in paraffin, and sections were stained with hematoxylin and eosin, and slides were examined by a board-certified veterinary pathologist. Dogs were enrolled if histopathology revealed intestinal inflammation with no identifiable underlying cause (such as infectious agents). Dogs were excluded if another histopathologic diagnosis was identified.

Three 14-day treatment tiers were included in the trial (FIG. 1A and Fig S1), and dogs were evaluated for a therapeutic response at the conclusion of each tier using CCECAI. Remission was determined using an abbreviated CCECAI that included scores to the first five indices (attitude/activity, appetite, vomiting, stool consistency, and stool frequency), and was defined as an abbreviated CCECAI score ≤2, with no score >1 for any of the five indices. Animals were first administered a therapeutic hydrolyzed protein diet (Royal Canin HP). Dogs that entered remission following this treatment were designated as diet-response (DR) and were maintained on therapeutic diet for the reminder of the trial. Animals that did not respond to therapeutic diet (NDR) subsequently began a two-week course of metronidazole (10 mg/kg PO q 12 hours) while being maintained on the therapeutic diet. Dogs that entered remission following antibiotic treatment were maintained on the combination of antibiotics and therapeutic diet for the reminder of the trial. Animals that still failed to show a favorable response remained on diet and metronidazole but received prednisone (1 mg/kg PO q 12 hours) (Tier 3) for the final 14 days of the trial. Dogs that presented with hypoalbuminemia (protein-losing enteropathy) at the initial screening were presumed to have more severe disease and poorer prognoses and thus were immediately administered all three interventions and were not included in the analyses. All dogs in which serum cobalamin was low at screening were supplemented with cyanocobalamin (50 mcg/kg SQ q 7 days) for the duration of the study. At the conclusion of the study, all animals returned to the clinic for the primary endpoint, which included a full re-evaluation of dogs including complete physical examination, complete blood count, serum chemistry, serum measurement of cobalamin and folate (if low at screening visit), urinalysis, CCECAI scoring and final fecal collections.

16S rRNA Gene Sequencing and Data Analysis

Genomic DNA was extracted from stool using the PowerSoil DNA Isolation Kit (MO BIO Laboratories, Carlsbad, Calif.) following the manufacturer's recommendations. A mock community pool containing purified genomic DNA from 12 known bacterial isolates was amplified and sequenced as a quality control. Additional controls included extraction of blank-processed samples (in which the DNA extraction process was followed without addition of input material), and water only, to determine background microbial signal. A dual-index amplicon sequencing method was employed for PCR amplification of the V4 region of the 16S rRNA gene⁶¹. Pico-green based Amplicons were sequenced on a MiSeq platform (Illumina, San Diego, Calif.) using 250 base pair paired-end chemistry. Reads were filtered to remove sequences with average Phred quality score ≤20 using Quantitative Insights into Microbial Ecology (QIIME)⁶² with filtering options (-q 20 -p 0.75 -r 3). Homopolymers >10 bp in length and sequences <248 bp and >255 bp were removed using Mothur⁶³. Chimeric sequences were identified and removed by usearch61⁶⁴ against the representative 16S sequences of SILVA128 (97_otus_16S.fasta)^(65,66). Quality-controlled sequences were then clustered against the SILVA128 database (SILVA_128_QIIME_release) using the open-reference OTU picking as implemented in QIIME with default parameters. The OTU table was rarefied to 10600 sequences per sample. In order to get a taxonomic assignment at species level for the OTUs from Clostridium sensu stricto 1, the corresponding representative sequences in SILVA database were used to search against NCBI ‘nr’ database. Species were temporarily assigned by the best hits (P<le-5) and further confirmation were done by comparing the relative abundances of these species determined by metagenomic shotgun sequencing method and by 16S sequencing method. The OTU ‘New.ReferenceOTU52’ represents C. perfringens, which is the most dominant OTU in some dogs, and the OTU ‘FJ957494.1.1454’ is corresponding to C. hiranonis.

Analysis of OTU tables was carried out using the R statistical environment⁶⁷, the bioconductor suite of software⁶⁸, and the Phyloseq2 package⁶⁹. Singletons and OTUs with ambiguous annotations were removed from the OTU table. Alpha diversity (Shannon diversity index and Faith's Phylogenetic Diversity) and Beta diversity (weighted and unweighted UniFrac) were calculated using Phyloseq2. Pielou's evenness index was calculated according to the literature⁷⁰. Functional potential of microbial communities (KEGG pathways and KEGG Orthologs) was predicted by Tax4Fun⁷¹ with default parameters against SILVA123 database. Wilcoxon sum rank test was used for comparisons of KEGG pathways at different timepoints (FDR <0.05). Principal component analysis for KEGG pathways and orthologs was performed by the R package factoextra. For differential abundance analysis and association analysis, filtering was carried out to remove taxa with a max abundance <0.1% across all samples and present in <10% of all samples. The resulting 381 species accounted for an average 96.23% of the total microbial composition. DESeq2⁷² implemented in Phyloseq2 (test=“Wald”, fitType=“parametric”) was used for differential abundance analysis on different taxonomy levels (Fold change >2 and P value <0.05) using un-rarefied reads. The Spearman correlation was computed between the abundance of each microbial composition (Log-transformed) and the values of different factors (i.e., CCECAI for each dog, time points, concentration of each metabolite). To avoid taking log of the zero value, 1 read was added to the abundance for each composition before calculating the Spearman correlation. All p values in the above analysis were adjusted by the FDR (Benjamini-Hochberg) method for multiple comparisons except where noted.

Metagenomic Sequencing and Data Analysis

Sequencing libraries were prepared using Illumina Nextera XT with 1 ng of canine stool collected at days 0, 14 and 42 from 19 out of the 20 diet-responsive dogs in the study. Sizing and quantification of libraries was carried out using a Tapestation 4200 (Agilent) and Qubit 3 (Thermo Fisher), respectively. Equimolar amounts of each library were pooled and sequenced on an Illumina NextSeq 500 instrument to produce 150 bp, paired-end sequences. Sequencing adapters and low quality reads were trimmed and filtered by Trimmomatic (v0.36) (leading:3 trailing:3 slidingwindow:4:15 minlen:36). High quality reads were mapped to the canine reference genome (CanFam3.1), using Bowtie2 v2.3.4.1 (--very-sensitive), and aligned reads were removed using SamTools⁷³. After host filtering, each sample was sequenced to a depth of >10 million paired-end reads (median depth=35.8 million). Taxonomic annotation for each sample was generated using Metaphlan2⁴⁶. The identified Clostridium spp. and Eubacterium spp. were further searched for the existence of genes involved in secondary bile acid production (bai operon) using tBlastn against the reference genomes of these species in GeneBank with the protein sequence of genes in 7α-dehydroxylation pathway (baiG, baiB, baiA, baiF, baiCD and baiE) (p-value ≤1e-5).

Metagenomic data from pediatric Crohn's disease patients before and after exclusive enteral nutrition (EEN) have been described previously²³ and were downloaded from European Nucleotide Archive (ENA) (SRP057027). The same filtering steps and settings for the metagenomic data analysis above in this study were used for these datasets. After filtering out human reads, taxonomic annotation for each sample using Metaphlan2 showed the presence of Clostridium. Among them, Clostridium scindens has been well known for the secondary BA producing ability. Paired reads with PCR duplicates removed by samtools⁷³ were aligned to the C. scindens reference genome (ASM15450v1, strain ATCC 35704) as well as strain VE202-05 (ASM47184v1) using bwa-mem (v0.7.17-r1188)⁷⁴ with default settings to estimate the abundances of bacteria among different samples (proportion of mapped reads in total reads). Wilcoxon sum rank test was used to test for significant differences in read mapping, and Spearman correlation was used to compare number of reads mapped with log-transformed fecal calprotectin (FCP) levels²³.

Anaerobic Culture and Identification of Bacterial Isolates by Whole Genome Sequencing

Rectal swabs freshly collected from dogs with active disease (day 0) and/or in remission at the end of the study (day 42) were transferred to an anaerobic chamber (97.5% nitrogen, 2.5% hydrogen; Coy Labs, Grass Lake, Mich.) within one hour of collection. The tip of the swabs was homogenized in 1 mL of pre-reduced PBS with 1% cysteine (PBSc). Serial dilutions made in PBSc (down to 10⁻⁵) were plated on brain-heart infusion (BHI), yeast casitone fatty acid with carbohydrate (YCFAC)⁷⁵, gut microbiota medium (GMM)⁷⁶, and De Man, Rogosa and Sharpe (MRS)⁷⁷ agars (Anaerobe Systems, Morgan Hill, Calif.). After incubation at 37° C. for 1-3 days, single colonies were picked from plates and grown overnight in BHI, YCFAC, GMM, or MRS broth (Anaerobe Systems, Morgan Hill, Calif.). Overnight cultures were saved as glycerol stocks (25% glycerol) and frozen neat for DNA extraction. DNA was purified from bacterial isolates using the High Pure PCR template kit (Roche) and used for PCR with primers specific for the bacterial 16S rRNA gene, including 27F (5′-AGAGTTTGATCMTGGCTCAG-3′), 515F (5′-GTGCCAGCMGCCGCGGTAA-3′), and 1492R (5′-CGGTTACCTTGTTACGACTT-3′). PCR products were purified using QiaQuick PCR Purification kit (Qiagen), Sanger sequenced, and sequences were assembled using Geneious software v11.1.5 (Biomatters Inc.). The longest high quality stretch of assembled sequence (at least 800 bp) was used for BLAST to find closest the match in Genbank. In addition, for selected C. hiranonis, C. perfringens and E. coli isolates, 1 ng of DNA was used to construct sequencing libraries using Illumina Nextera XT. Libraries were sized and quantified as described above for metagenomic sequencing. For each sample, at least 10 million, 150 bp single-end reads were generated using an Illumina NextSeq 500 instrument. Quality control steps were the same as the metagenomic analysis above. High quality reads were mapped to the genome of C. hiranonis (ASM15605v1) using Stampy⁷⁸ (--substitutionrate=0.1), which allows mapping of reads that are highly divergent from the reference genome. PCR duplicates were removed by Samtools. Coverage of genomic regions representing the bai operon were calculated for each isolate to show the existence of genes in 7α-dehydroxylation pathway.

Metabolomics and In Vitro Bacterial Growth Inhibition Assays

Bile acids were quantified in stool using a Waters Acquity uPLC System with a QDa single quadrupole mass detector and an autosampler (192 sample capacity) as described previously⁷⁹. Briefly, fecal samples were suspended in methanol (5 μL/mg stool), vortexed for 1 minute, and centrifuged at 13,000 g for 5 minutes. The supernatant was transferred to a new vial and analyzed on an Acquity uPLC with a Cortecs UPLC C-18+1.6 mm 2.1×50 mm column. All chemicals and reagents were mass spectrometry grade. Canine isolates of C. perfringens (n=3) and E. coli were revived from glycerol stocks in Modified Reinforced Clostridial Broth (MRCB, Fisher Scientific) or Luria broth (LB, Fisher Scientific), respectively and grown overnight in the anaerobic chamber at 37° C. Lithocholic and deoxycholic acids (Sigma) were dissolved in 100% ethanol (30 mg/mL). Growth inhibition by deoxycholic acid was determined by microbroth dilution and assessed by OD 630 after overnight growth. Due to low solubility (<1 mg/L), inhibition by lithocholic acid was assessed by counting colonies on agar plates with LCA (0, 0.1, 0.25, 0.5, 0.75, or 1 mg/mL and LB plates for E. coli, and 0, 0.01, 0.025, 0.05, 0.075, or 0.01 mg/mL and Columbia blood agar supplemented with 5% defibrinated sheep's blood for C. perfringens that were incubated anaerobically, at 37° C. for 24 (E. coli) or 48 (C. perfringens) hours.

Mouse Experiments

Female C57BL/6 (7 weeks old) (Jackson Laboratory) were orally pre-colonized with a kanamycin-resistant E. coli strain (Nissle 1917) (1×10⁹ CFU/mouse) 4 days prior to the to the start of dextran sulfate sodium (DSS) treatment. Animals were randomly assigned to groups (cages) at baseline and drinking water was replaced with either filter-sterilized water (mock-treatment), or a filter-sterilized solution of 2.5% (w/v) DSS (relative molecular mass 40,000; Sigma-Aldrich) in water. The mice treated with mock or DSS were orally gavaged C. hiranonis (1×10⁸ CFU/mouse, in anaerobic PBS) or PBS (control) from days 0 to 4. C. hiranonis was grown overnight in MRCB, anaerobically, at 37° C. Culture density was assessed via optical density (630 nm) and the required volume of culture was spun at 10,000 g for 15 min. Bacterial pellets were resuspended in PBS to obtain a dose of 1×10⁸ CFU/mouse. All procedures were performed in accordance with the guidelines of the University of Pennsylvania Institutional Animal Care and Use Committee. The mice were then euthanized at day eight and colon contents and tissues were collected. Colon contents were weighted and cultured on LB agar plates with kanamycin (100 μg/mL) for 16 hours. Stool samples from baseline and colon contents from day eight were collected and stored at −80° C. for the detection of bile acid levels. Colons were fixed in formalin and sections stained with haematoxylin and eosin (H&E). Pathology was blindly evaluated by an board-certified veterinary pathologist (C.B.) according to standard criteria for DSS colitis.

Data Availability

Raw 16S rRNA gene sequences for canine stool samples have been deposited in the Sequence Read Archive (SRA so; accession number pending). Processed OTU tables and metadata can be accessed through MicrobiomeDB⁵⁶. Metagenomic and whole genome sequence data are also available on SRA (accession numbers pending).

Results Dietary Therapy Induces Rapid and Durable Remission

To investigate the impact of a therapeutic diet on disease and the microbiome, treatment-naive dogs (n=29) with chronic enteritis (CE) were enrolled in a study to evaluate the impact of diet on disease and the microbiome. Dogs with active disease were switched from their current diet to a commercially-available therapeutic hydrolyzed protein diet (FIG. 1A). Impact of treatment on disease was monitored using the Canine Chronic Enteropathy Clinical Activity Index (CCECAI; hereafter referred to as ‘disease score’), which is positively correlated with poor clinical outcome³⁶. After two weeks on therapeutic diet, 69% ( 20/29) of animals entered remission, marked by a reduction in the mean disease score from 4.1 (95% CI=4.8-3.3) to 1.3 (95% CI=1.8-0.7). These diet-responsive (DR) animals were maintained on diet for the remainder of the study with no additional interventions (FIG. 1B). At the conclusion of the study (day 42), DR animals had an mean disease score of 0.9 (95% CI=1.3-0.6), constituting an >4-fold reduction in disease severity compared to day 0 (FIG. 1B). In contrast, 31% ( 9/29) of animals failed to show a significant reduction in disease score after two weeks on therapeutic diet (FIG. 1C). These non-diet-responsive (NDR) animals presented with more severe disease scores (mean score=6.1; 95% CI=7.4-4.7) than DR animals (P<0.05 at day 0) and did not show a significant reduction after 2 week diet therapy (FIG. 1C). NDR animals were maintained on diet therapy for the reminder of the study, but also received combination therapy that included antibiotics (at day 14) and prednisone (at day 28) (FIG. 1A and FIG. 8, see methods), but showed only incremental improvement in disease scores (FIG. 1C). These data highlight a rapid clinical response to hydrolyzed diet in the majority of dogs with chronic enteritis.

Identification of Microbial Community Profiles Associated with Treatment Outcome

To determine whether treatment with hydrolyzed diet alone is sufficient to alter the microbial community in the gut, 16S rRNA gene profiling was carried out on fecal samples collected from DR, NDR and healthy control animals (n=11). Consistent with previous reports³⁸, it was found that the diversity of the canine fecal microbiome was not significantly altered in dogs with CE, compared to healthy controls (FIGS. 9A-B), and that the communities in both groups were predominantly comprised of Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, and Fusobacteria (FIG. 9C). However, compared to healthy dogs, animals with CE showed greater between-individual distance in microbial community structure by unweighted Unifrac (FIG. 9D). Using a ternary plot visualization, an enrichment of Operational Taxonomic Units (OTUs) was observed from Firmicutes and Proteobacteria in animals with active disease, while Bacteroidetes were enriched in healthy animals (FIG. 2A). Interestingly, a subset of proteobacterial OTUs was highly enriched in DR animals compared to both NDR and healthy controls (FIG. 2A), tan points in lower left corner).

These differences prompted us to carry out a formal differential abundance analysis, identifying 55 OTUs that distinguish healthy animals from those with disease (Table 2). For example, Escherichia coli, which is commonly associated with intestinal diseases, was over-represented in animals with CE (FIG. 2B), showing a significant, albeit weak, positive correlation with disease score (R=0.2109, P=0.02626) (FIG. 2C). OTUs from Clostridium sensu stricto 1 were also enriched in CE, including Clostridium perfringens (FIG. 2D), which was also positively correlated with disease scores (FIG. 2E) (R=0.2324, P=0.01412). These bacteria have been implicated in large bowel diarrhea/colitis in dogs³⁹. Taken together with previously published work⁴⁰, these data demonstrate that dysbiosis during CE is marked by the presence of pathobionts. Next, whether the microbiome in DR and NDR animals differed prior to the start of treatment (day 0) was investigated. Although no differences were observed between the two groups in community diversity, evenness or distance from healthy controls (unweighted or weighted Unifrac), 21 OTUs were identified that were differentially abundant between DR and NDR animals, 13 of which were enriched in animals that ended up responding to diet treatment (FIG. 2F and Table 3). Interestingly, Proteobacteria and C. perfringens were found to be more abundant in DR animals (FIG. 2F). Collectively, these results highlight distinct microbial signatures during disease that are associated with different clinical outcomes following diet therapy.

Therapeutic Diet Ameliorates Dysbiosis Associated with Chronic Enteritis

To assess whether diet-induced remission is accompanied by alterations in dysbiosis, the microbial community structures were compared before and after administration of therapeutic diet in DR animals. No significant change was observed in phylogenetic distance or shannon diversity (FIG. 10A-B) but did see a marked increase in community evenness following diet administration (FIG. 3A) when focusing on the top 40 most abundant OTUs among the samples, which account for 83% of the total reads. Principal coordinate analysis based on unweighted (FIG. 3B) or weighted (FIG. 10C) UniFrac showed a clear separation between dogs, even at day 0, before diet therapy was administered, highlighting heterogeneity in dysbiosis associated with clinical disease. Despite this baseline difference between animals, community structure underwent a marked shift away from disease-state by 14 and 42 days after diet therapy (FIG. 3B). Comparing unweighted Unifrac distances between DR and healthy animals at each time point, it was observed that diet-induced remission was marked by decreased phylogenetic distance relative to healthy controls, a trend that continued through day 42, when the phylogenetic similarity to day 0 was lowest and similarity to healthy dogs was highest (FIG. 3C).

Given that therapeutic diet shifted the community structure of the microbiome in DR animals, it was reasoned that composition of the fecal microbiome would be rapidly altered by dietary intervention. Administration of therapeutic diet was broadly characterized by an increase of Firmicutes and a decrease of Proteobacteria (FIG. 3D). Fourteen days after beginning diet therapy, ten genera were differentially abundant compared to pre-treatment (day 0) in DR animals (Table 4). Potential pathogenic genera associated with IBD were found under-represented after diet treatment. For example, Escherichia-Shigella, Clostridium sensu stricto 1, Enterococcus and Fusobacterium had a higher relative abundance at Day 0 and were significantly reduced after 14 days on therapeutic diet. When evaluated at species level, 36 OTUs were significantly differential abundant between samples collected at day 0 compared to day 14 (FIG. 3E) (Table 5). E. coli was typically enriched in the animals at day 0 in this study (FIG. 3E), and its relative abundance declined dramatically after two-weeks on therapeutic diet, eventually reaching levels nearly undetectable by day 42 that were also indistinguishable from levels observed in healthy dogs (FIG. 3F). C. perfringens also showed significant lower prevalence in the samples at day 14 and in healthy dogs, compared to day 0 samples (FIG. 3G). In turn, several increased OTUs were from the genera that have been suggested as beneficial commensals in human studies, such as Blautia⁴¹ (Table 5). Taken together, these results point to ameliorated dysbiosis with a reduction of pathobionts and increase of beneficial commensal taxa as a hallmark of diet therapy.

Remission-Specific Changes in the Microbiome Following Diet Therapy

It was hypothesized that the changes observed following diet therapy in DR animals are associated with remission, rather than merely a response to diet that is independent of clinical outcome. To test this hypothesis, the impact of therapeutic diet on dogs that entered remission following diet therapy alone (DR), was compared with changes observed in non-diet-responsive (NDR) animals that failed to enter remission after diet therapy, and which require additional therapies after day 14. Whereas diet therapy in DR animals was associated with increased community evenness (FIG. 3A) and a decreased phylogenetic distance from health dogs (FIG. 3C), the same treatment in NDR dogs did not significantly affect the microbial community evenness or Unifrac distance to healthy dogs (FIGS. 4A and 4B). Just as it was observed in DR animals (FIG. 3D), diet also altered the gut microbiota compositions in NDR animals (FIG. 4C). Differential abundance analysis comparing NDR animals at day 0 versus day 14, when they received only therapeutic diet, identified 24 OTUs (Table 6). However, this shift was distinct from that observed in DR animals (FIG. 4D and FIG. 11). For example, diet therapy was associated with a decrease in the relative abundance of Fusobacterium and Phascolarctobacterium in NDR animals at day 14, compared to day 0, while these taxa were either unchanged or more modestly altered by diet therapy in DR animals. Conversely, Escherichia-Shigella, Enterococcus and some of Clostridium sensu stricto 1 are only reduced in animals that enter remission after diet treatment (FIG. 4D). The disease associated bacteria E. coli and C. perfringens were not significantly changed in NDR animals after diet therapy (FIGS. 4E and 4F). After 14 days on therapeutic diet, NDR dogs were maintained on diet, but were also administered metronidazole, an antibiotic that largely targets anaerobes. Interestingly, antibiotic treatment exacerbated dysbiosis, resulting in a precipitous decline in community evenness (FIG. 4A), increased distance from healthy controls (FIG. 4B), and increased relative abundance of potential pathogens (FIGS. 4E and 4F).

Diet-Induced Remission is Associated with Metabolic Reprogramming and Increased Levels of Secondary Bile Acids.

To determine if diet induced changes in microbial community structure would translate to altered microbial metabolism the 16S rRNA gene sequencing data were used to assess the relative abundance of predicted KEGG pathways. Principal component analysis of metabolic pathway abundance data showed a separation between samples from animals with active disease (day 0) and those in remission (day 14) (FIG. 12, FIGS. 5A and 5B). Differential abundance analysis identified 36 pathways were increased in relative abundance as a result of diet treatment in DR animals (Table 7), including several involved in carbohydrate metabolism and secondary bile acid synthesis (FIGS. 5C and 5D). In contrast, 50 pathways were reduced, including fatty acid and steroid biosynthesis (FIG. 5C). This shift in metabolic potential away from lipid biosynthesis toward carbohydrate and bile acid synthesis as animals entered remission prompted us to quantify bile acids in stool. Using targeted metabolomics, levels of 15 bile acids in stool of healthy dogs were measured, compared with stool collected at days 0, 14 and 42 in the study (FIG. 13, Table 10). Consistent with the 16S data, the secondary bile acids deoxycholic acid (FIG. 5E) and lithocholic acid (FIG. 5F) were high in healthy controls but low in animals with active disease (day 0) and were significantly increased after the diet treatment in DR animals at day 14 and 42 (FIGS. 5E and 5F, and Table 8). Notably, levels of these secondary bile acids were not elevated by diet treatment in NDR animals (FIGS. 5G and 511), suggesting that this metabolic shift is linked to diet-induced remission.

Lithocholic and Deoxycholic Acid Inhibit the Growth of E. coli and C. perfringens, In Vitro.

Diet-induced remodeling of the microbiome could be due, at least in part, to the inhibitory role of secondary bile acids on harmful bacteria. Correlation analysis of the metabolomics and microbiome data identified thirteen genera significantly associated with at least one bile acid (Spearman, R >0.04 or <−0.04) (FIG. 6A). The primary bile acid, cholic acid, was negatively correlated with 11 OTUs, consistent with the reported ability of this bile acid to negatively regulate bacterial growth⁴². It was also observed that the increase in secondary bile acids following diet treatment correlated with a reduction in relative abundance of certain bacteria (e.g., OTUs from Escherichia-Shigella, Clostridium and Fusobacterium) (Table 9). To directly test this hypothesis, lithocholic or deoxycholic acid were assessed for their ability to inhibit the in vitro growth of E. coli (n=1) or C. perfringens (n=3) isolates derived from the dogs with active disease, since these species or their genera were associated with disease in the animal model. Deoxycholic acid blocked the growth of both species at a concentration comparable to what was detected in the fecal samples (FIGS. 6C and 6E), while lithocholic acid blocked the growth of E. coli but not C. perfringens (FIGS. 6B and 6D, respectively). Collectively, these results show that the inhibitory activity of these bile acids varies for different bacteria and suggest that elevated secondary bile acids observed following diet therapy can contribute to the decrease of potentially harmful bacteria. C. hiranonis is a diet-responsive species with the ability to produce secondary bile acids.

Next, the source of lithocholic and deoxycholic acids after diet treatment was identified. Production of these from primary bile acids requires the 7-dehydroxylation activity conferred by the bile acid-inducible (bai) operon—an activity unique to a limited number of anaerobes representing a small fraction of the microbiome, including some Clostridial and Eubacterial species⁴³ Given the finding that certain clostridial OTUs, as well as levels of lithocholic and deoxycholic acids, increase after diet-induced remission (FIG. 3G and FIG. 5E-F, respectively), potential bile acid producers in DR animals were identified. Stool samples collected day 0, 14 and 42 after starting diet therapy were subjected to metagenomic sequencing. Taxonomic assignment of reads identified six Clostridium species (C. perfringens, C. hiranonis, C. nexile, C. colicanis, C. glycolicum and C. ramosum) and 2 Eubacterium species (Eubacterium biforme and E. dolichum) present in these samples at a relative abundance ≥0.01% in at least 10% samples. Of these species only C. hiranonis has been reported to have the bai operon⁴⁴, and this was confirmed by BLAST against the reference genomes of these species in GenBank. Moreover, the metagenomic data (FIG. 14) and 16S sequencing data showed that the relative abundance of C. hiranonis was significantly increased after diet treatment in DR animals (FIG. 6F, left) but not in NDR animals that failed diet therapy (FIG. 6F, right). Since the Clostridium genus exhibits a high level of genetic divergence, even at the species level, that canine C. hiranonis possesses the bai operon was confirmed. Anaerobic culture of rectal swabs collected during the study, followed by isolate picking and Sanger sequencing of full-length 16S rRNA gene, was used to assemble a canine culture collection from 7 dogs with chronic enteritis before and/or after treatment. In total, 49 Clostridium isolates belonging to 5 species were identified (C. baratii, C. perfringens, C. sartagoforme, C. hiranonis, and C. lactatifermentans). 82% ( 31/39) of the clostridial isolates from animals with active disease were C. perfringens, consistent with the reported involvement of this organism in canine³⁹ and human⁴⁵ gastrointestinal disease. Two C. hiranonis isolates were obtained from independent diet-responsive animals in remission at day 42. These C. hiranonis isolates and three C. perfringens isolates were selected for whole genome sequencing. Reads were aligned to the reference C. hiranoni, revealing an intact bai operon in canine C. hiranonis, but not C. perfringens (FIG. 6G). Taken together, these data point to C. hiranonis, a species originally isolated from human stool⁴⁴, as a likely bile acid producer associated with diet-induced remission in dogs.

C. hiranonis Inhibits Inflammation-Induced Expansion of E. coli in a Mouse Model of DSS Colitis

The ability of C. hiranonis to produce secondary bile acids, combined with the observation that lithocholic and deoxycholic acids were potent inhibitors of E. coli and C. perfringens growth, in vitro, prompted us to test whether C. hiranonis could restrict expansion of potential pathogens in vivo during inflammation. Mice were first colonized with drug-selectable E. coli (Nissle 1917 strain), inflammation was triggered by administration of dextran sodium sulfate (DSS) in the drinking water, and animals were either orally administered PBS daily (mock) or C. hiranonis (FIG. 611). DSS treatment resulted in reduced colon length (FIG. 6I), and a dramatic bloom of the E. coli Nissle strain (FIG. 6J). In contrast, DSS-treated mice that received C. hiranonis daily showed a marked reduction of colonic shortening, and a near complete abrogation of E. coli expansion. Taken together with the finding that lithocholic and deoxycholic acids can inhibit growth of pathobionts, these data suggest that C. hiranonis or secondary bile acids produced by this species, mediate colonization resistance during enteritis.

C. scindens is Associated with Diet-Induced Remission in Pediatric Crohn's Disease

Given that high remission rates are observed in both dogs and humans following dietary therapy, it was hypothesized that a similar induction of bai operon-containing clostridia can occur in pediatric Crohn's disease patients being treated with exclusive enteral nutrition (EEN). To test this, publicly available data from a recent study that examined approximately 20 patients before and after treatment with EEN were analyzed²³, in which half responded to treatment while other half failed EEN therapy. Classifications of bacterial taxa present in each sample using standard metagenomic analysis methods⁴⁶ revealed the presence of C. scindens, which is recognized for having high 7-dehydroxylation activity^(44,47). The relative abundance was further estimated using the proportion of total reads that map the reference genome of C. scindens. As shown in FIG. 7A, this bacterium increased significantly from pretreatment to 8 weeks post-EEN, as did the number of reads mapping to the bai operon (FIG. 7B). Remarkably, this increase was only observed in patients that entered remission following EEN (Responsive, n=10) but not those that failed therapy (Non.Responsive, n=10) (FIGS. 7A and 7B). In addition, the correlation analysis between the relative abundance of C. scindens and fecal calprotectin (FCP), a biomarker of disease activity for IBD²³, indicated a significantly negative correlation (FIG. 7C) in EEN ‘Responsive’ patients (R=−0.3515, P=0.03287), but not in EEN ‘Non.Responsive’ patients (R=−0.0267, P=0.877). Similarly, a significant negative correlation between bai operon and FCP was observed in diet-responsive (R=−0.3944, P=0.0157), but not non-responsive patients (R=0.0490, P=0.7766) (FIG. 7D). These results collectively point to bile acid producing clostridia as key features of diet-induced remission and potent inhibitors of pathobiont colonization in both animals and humans (FIG. 7E).

Discussion

Using a diet-responsive animal model to study the role of the microbiome in chronic enteritis, remission-specific changes in microbiome composition and function were identified. All animals enrolled in the study had active disease, yet their baseline microbiome composition differed greatly (FIG. 3), perhaps reflecting variation in their environment, genetic background (breed), age and weight. This variation in composition supports the idea that enteric disease is driven not by a single dysbiotic state, but rather dysbiosis reflects a loss of community stability⁴⁸. Rather than dramatic changes in microbial community structure following diet therapy, a shift from lipid metabolism to carbohydrate and bile acid synthesis was observed. Although the metabolomics analysis was focused on bile acids, a broader picture of the metabolites produced before and after diet therapy, as well as the macro- and micro-nutrients present in the diets themselves, can improve the understanding of the mechanisms by which diet achieves remission.

One important open question is precisely how therapeutic diets such as EEN or prescription pet foods alter the microbiome and whether there are general principles that could be used to guide the development of better dietary therapies. Studies in pediatric Crohn's disease have reported higher remission rates with EEN compared to partial enteral nutrition (PEN), which includes some table food. These observations have led some to postulate that a highly monotonous diet that is reduced in complexity can constitute an essential part of nutritional therapies for IBD. Consistent with this notion, mice fed a monotonous diet exhibited lower microbial diversity and were more susceptible to DSS colitis than mice fed an alternating diet⁴⁹. However, the prevalence and treatment of chronic enteritis in veterinary medicine highlights that disease routinely develops even when diets are monotonous and that rapid and robust remission can be achieved with solid food. Hydrolyzed protein diets, such as the one used in the study, have been shown to be effective in the management of canine chronic enteropathies^(50,51), and have previously been shown to be more effective for long term management when compared to a highly digestible diet formulated with non-hydrolyzed protein sources^(50,51) While it is uncertain what characteristic of these hydrolyzed formulas are driving the response, the low molecular weight of hydrolyzed proteins can reduce their ability to be recognized by the immune system while providing improved digestibility. In summary, the results suggest that the dog would be a useful model to dissect the beneficial and harmful roles of different diets, particularly since formulated diets have long been a standard of care for treating numerous diseases in companion animals.

Secondary bile acids and bile acid-producing clostridial species were identified as key features of diet-induced remission in humans and dogs. These findings complement recent studies examining the mechanisms by which fecal microbiota transplant (FMT) cure Clostridium difficile infection⁵². Buffy et al. identified Clostridium scindens as associated with resistance to C. difficile infection in both humans and mice, and they showed that transfer of C. scindens, or a consortium containing this organism, protected mice from C. difficile challenge. Moreover, inhibition of C. difficile growth in vitro by C. scindens was associated with secondary bile acid production. These data are consistent with microbiological studies showing that primary bile acids induce germination of C. difficile, while certain secondary bile acids can block vegetative growth⁵³. Although C. difficile was not observed in the animals, C. perfringens and E. coli were identified as major disease-associated taxa, and it was shown that physiologic levels of secondary bile acids potently block in vitro growth of these organisms. It is not known whether bile acids can restrict these organisms in vivo in the canine model but elucidating these mechanisms could have important health implications beyond veterinary medicine. Although C. difficile is the leading cause of nosocomial diarrhea in humans, C. perfringens and E. coli are both common human commensals and have been implicated in both diarrheal disease and colitis in humans and dogs. Moreover, the ability of C. perfringens to produce numerous toxins make it a leading cause of foodborne illness and soft tissue infections. Interestingly, when data from a cohort of pediatric Crohn's disease patients before and after diet therapy were examined, it was found that C. scindens was associated with diet-induced remission (FIG. 7), and a related study showed that sustained remission following EEN was characterized by low levels of proteobacteria, while patients that relapsed showed a marked increase in proteobacteria⁵⁴. The data, together with previous studies in dogs^(33,35), highlight the importance of leveraging animal models and advocate for the use of newly developed analytical methods⁵⁵ and database approaches^(56,57) for comparing across multiple microbiome studies to take a ‘One Health’ approach that could identify conserved themes in host-microbiome interactions.

C. difficile infections frequently arise after antibiotic treatment, a phenomenon attributed to the effect of antibiotics on secondary bile acid levels⁵⁸. Interestingly, it was also observed that antibiotics antagonized the diet-induced shifts in microbiome composition and function, promoting a more dysbiotic state coincident with dramatically reduced levels of lithocholic and deoxycholic acid (FIG. 4 and FIG. 5). Taken together, these data support a more general model for microbe-microbe interactions in the gut in which bile acid producing clostridia restrict the growth of a range of bile acid-sensitive pathobionts to limit disease and highlight that these processes are exquisitely sensitive to antimicrobials. The parallels between the findings and those reported for FMT and C. scindens would suggest that FMT might also be beneficial for treating enteritis. Clinical trials testing this hypothesis in IBD patients have shown moderate success, in marked contrast to C. difficile infections where FMT is curative for the vast majority of patients⁵⁹. This discrepancy can be related to different pathobionts contributing to IBD pathogenesis. Interestingly, colitis is a common side-effect observed in cancer patients undergoing immune checkpoint blockade, and a recent study demonstrated complete resolution of this colitis following FMT⁶⁰, raising the possibility that bile acid producers can be important in treating certain types of colitis.

TABLE 2 OTUs with differential abundances between samples of healthy dogs and dogs with CCE at day 0 (Fold change >2 and P-value <0.05). log2- Base Fold- P OTU Mean Change value Kingdom Phylum Class Order Family Genus Species HQ802983.1.1440 134.78 −9.14 3.28E−10 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Tyzzerella 4 NA FJ950694.1.1472 2052.29 −5.56 6.45E−09 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella Escherichia coli HG798451.1.1400 30.74 −6.70 1.14E−07 Bacteria Firmicutes Bacilli Lactobacillales Enterococcaceae Enterococcus Enterococcus durans New.ReferenceOTU52 676.55 −6.51 8.89E−07 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium sensu stricto 1 perfringens New.ReferenceOTU82 35.49 −4.87 6.27E−05 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium sensu stricto 1 NA GQ449092.1.1375 69.30 −7.04 7.97E−05 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Tyzzerella NA FJ506371.1.1371 34.88 −5.40 0.00012 Bacteria Firmicutes Erysipelotrichia Erysipelotrichales Erysipelotrichaceae Erysipelatoclostridium NA GQ448744.1.1393 81.06 −6.86 0.00011 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides Ambiguous_taxa FJ957494.1.1454 19.18 −6.33 0.00037 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium sensu stricto 1 Ambiguous_taxa HQ760911.1.1437 11.66 −5.91 0.00045 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Anaerostipes NA GQ006324.1.1342 10.73 −5.77 0.00084 Bacteria Actinobacteria Actinobacteria Corynebacteriales Corynebacteriaceae Corynebacterium 1 uncultured bacterium GQ448246.1.1389 313.30 −3.87 0.00079 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides Ambiguous_taxa KC245406.1.1465 3.79 −3.70 0.0007 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Lachnoclostridium uncultured bacterium New.ReferenceOTU54 39.95 −5.91 0.0009 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium HQ751549.1.1448 10.41 −4.70 0.0012 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae uncultured NA JF712675.1.1540 6.38 −5.03 0.0012 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella uncultured bacterium JQ208181.1.1352 148.49 −2.99 0.0012 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae [Ruminococcus] Ambiguous_taxa gauvreauii group GX182404.8.1529 3.29 −4.08 0.0020 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella NA FP929060.3837.5503 375.58 −1.82 0.0020 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae NA NA FN667392.1.1495 12.19 −5.97 0.0025 Bacteria Firmicutes Bacilli Lactobacillales Lactobacillaceae Lactobacillus uncultured bacterium FN667422.1.1495 5.84 −4.33 0.0034 Bacteria Firmicutes Bacilli Lactobacillales Lactobacillaceae Lactobacillus Ambiguous_taxa HK557089.3.1395 1340.69 −3.02 0.0046 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium HQ803964.1.1435 335.70 −2.90 0.0046 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Lachnoclostridium uncultured bacterium AM276759.1.1484 6.84 −2.87 0.0045 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium HK555938.1.1357 21.72 −6.40 0.0054 Bacteria Actinobacteria Coriobacteriia Coriobacteriales Coriobacteriaceae Collinsella uncultured bacterium KF842598.1.1394 22.60 −6.80 0.0054 Bacteria Bacteroidetes Bacteroidia Bacteroidales Porphyromonadaceae Parabacteroides NA HQ792778.1.1436 5.38 3.62 0.0058 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides uncultured FM865905.1.1392 8.52 −5.45 0.0066 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium NA sensu stricto 1 FN563300.1.1447 1147.14 −1.92 0.0064 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium HQ754680.1.1441 10.15 −2.20 0.0065 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae NA NA GQ867426.1.1494 3.36 −4.08 0.0072 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella uncultured bacterium EU470512.1.1400 2.07 −3.40 0.0079 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella uncultured bacterium AY239462.1.1500 2.71 −3.20 0.0080 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae [Ruminococcus] Ambiguous_taxa gauvreauii group New.ReferenceOTU114 8.57 −3.10 0.0091 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium FN668375.4306350.4307737 9.14 −4.09 0.0093 Bacteria Firmicutes Clostridia Clostridiales Peptostreptococcaceae NA NA AB009242.1.1451 8.33 −4.81 0.0097 Bacteria Spirochaetae Spirochaetes Spirochaetales Spirochaetaceae Treponema 2 NA HQ792787.1.1438 1.61 3.45 0.0128 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides uncultured bacterium AB506370.1.1516 5.92 −4.63 0.0194 Bacteria Bacteroidetes Bacteroidia Bacteroidales Prevotellaceae Prevotellaceae UCG-001 Ambiguous_taxa DQ057365.1.1393 5.11 −4.42 0.0202 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Lachnoclostridium Ambiguous_taxa FN667084.1.1493 8.26 −3.53 0.0216 Bacteria Firmicutes Bacilli Lactobacillales Lactobacillaceae Lactobacillus uncultured bacterium DQ113765.1.1450 1500.29 3.82 0.0230 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides NA HK694029.9.1487 6.57 −2.66 0.0244 Bacteria Firmicutes Erysipelotrichia Erysipelotrichales Erysipelotrichaceae Faecalitalea NA AJ270486.1.1241 10.92 −4.24 0.0290 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Coprococcus 1 Ambiguous_taxa EU768569.1.1352 5.69 −3.37 0.0314 Bacteria Firmicutes Clostridia Clostridiales Ruminococcaceae Ruminiclostridium 9 uncultured bacterium FM179752.1.1686 1.66 −3.11 0.0324 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Pseudocitrobacter NA JF807116.1.1260 2.54 −3.70 0.0351 Archaea Euryarchaeota Methanobacteria Methanobacteriales Methanobacteriaceae Methanobrevibacter uncultured archaeon FJ957528.1.1445 14.75 −5.09 0.0356 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Sarcina uncultured bacterium KC504009.1.1465 3.67 −3.15 0.0350 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella NA GQ448506.1.1374 304.58 −1.58 0.0335 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium JF224013.1.1362 2.89 −3.89 0.0390 Bacteria Bacteroidetes Bacteroidia Bacteroidales Porphyromonadaceae Porphyromonas uncultured bacterium EU774020.1.1361 12.85 2.51 0.0391 Bacteria Fusobacteria Fusobacteriia Fusobacteriales Fusobacteriaceae Fusobacterium uncultured bacterium GQ448486.1.1387 48.95 −2.72 0.0384 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium HQ793763.1.1451 13.41 3.32 0.0434 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides NA JN387556.1.1324 164.12 −2.78 0.0459 Bacteria Firmicutes Clostridia Clostridiales Peptostreptococcaceae Intestinibacter NA New.ReferenceOTU109 34.47 3.54 0.0488 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides NA

TABLE 3 OTUs with differential abundances between day 0-samples of diet responsive dogs and diet non-responsive dogs. Base log2Fold- OTU ID Mean Change P value Kingdom Phylum Class JRPJ01000002.1034290.1035971 111.72 8.05 0.000330185 Bacteria Proteo- Epsilon- bacteria proteobacteria JF920309.1.1340 22.79 5.56 0.00409 Bacteria Proteo- Epsilon- bacteria proteobacteria FJ978526.1.1378 8.93 5.36 0.04034 Bacteria Proteo- Gamma- bacteria proteobacteria New.ReferenceOTU45 47.39 23.74 3.66E−14 Bacteria Proteo- Gamma- bacteria proteobacteria HK555938.1.1357 19.48 −6.16 0.04481 Bacteria Actino- Coriobacteriia bacteria FJ957494.1.1454 20.74 −3.37 0.01429 Bacteria Firmicutes Clostridia New.ReferenceOTU52 1115.31 3.54 0.01242 Bacteria Firmicutes Clostridia DQ797046.1.1403 14.74 4.64 0.02361 Bacteria Firmicutes Negativicutes GQ449092.1.1375 15.00 −3.58 0.04561 Bacteria Firmicutes Clostridia AMCI01001631.34.1456 77.88 −4.66 0.00349 Bacteria Bacteroidetes Bacteroidia KF842598.1.1394 6.52 5.98 0.02931 Bacteria Bacteroidetes Bacteroidia HQ793763.1.1451 5.57 5.09 0.01396 Bacteria Bacteroidetes Bacteroidia DQ113765.1.1450 201.87 5.02 0.00453 Bacteria Bacteroidetes Bacteroidia ACBW01000012.3536.5054 4.37 4.18 0.02909 Bacteria Bacteroidetes Bacteroidia HK693629.1.1491 23.00 −2.59 0.01037 Bacteria Firmicutes Clostridia JQ208053.1.1336 14.50 2.58 0.04454 Bacteria Fusobacteria Fusobacteria GQ493166.1.1359 231.28 −2.75 0.00391 Bacteria Firmicutes Clostridia GQ448486.1.1387 39.63 −4.02 0.00023 Bacteria Firmicutes Clostridia GQ491426.1.1332 461.01 −2.89 0.03894 Bacteria Firmicutes Clostridia New.ReferenceOTU54 18.10 5.21 0.00152 Bacteria Firmicutes Clostridia JN387556.1.1324 167.47 3.99 0.00791 Bacteria Firmicutes Clostridia OTU ID Order Family Genus Species JRPJ01000002.1034290.1035971 Campylo- Helico- Helicobacter Ambiguous_taxa bacterales bacteraceae JF920309.1.1340 Campylo- Campylo- Campylo- NA bacterales bacteraceae bacter FJ978526.1.1378 Aero- Succinivi- Succinivibrio uncultured monadales brionaceae bacterium New.ReferenceOTU45 Aero- Succinivi- Anaero- Ambiguous_taxa monadales brionaceae biospirillum HK555938.1.1357 Corio- Corio- Collinsella uncultured bacteriales bacteriaceae bacterium FJ957494.1.1454 Clostridiales Clostridiaceae 1 Clostridium Ambiguous_taxa sensu stricto 1 New.ReferenceOTU52 Clostridiales Clostridiaceae 1 Clostridium NA sensu stricto 1 DQ797046.1.1403 Seleno- Veillonellaceae Allisonella uncultured monadales bacterium GQ449092.1.1375 Clostridiales Lachnospiraceae Tyzzerella NA AMCI01001631.34.1456 Bacteroidales Bacteroidaceae Bacteroides uncultured bacterium KF842598.1.1394 Bacteroidales Porphyro- Para- NA monadaceae bacteroides HQ793763.1.1451 Bacteroidales Bacteroidaceae Bacteroides NA DQ113765.1.1450 Bacteroidales Bacteroidaceae Bacteroides NA ACBW01000012.3536.5054 Bacteroidales Bacteroidaceae Bacteroides uncultured bacterium HK693629.1.1491 Clostridiales Lachnospiraceae Blautia NA JQ208053.1.1336 Fuso- Fuso- Fuso- NA bacteriales bacteriaceae bacterium GQ493166.1.1359 Clostridiales Lachnospiraceae NA NA GQ448486.1.1387 Clostridiales Lachnospiraceae Blautia uncultured bacterium GQ491426.1.1332 Clostridiales Lachnospiraceae Blautia uncultured bacterium New.ReferenceOTU54 Clostridiales Lachnospiraceae Blautia uncultured bacterium JN387556.1.1324 Clostridiales Peptostrepto- Intestinibacter NA coccaceae

TABLE 4 Genera with differential abundances between samples of dogs at day 14 and day 0 (day 14 versus day 0) for diet responsive dogs. Base log2Fold- P OUT IDs Mean Change value Kingdom Phylum Class Order Family Genus GQ006324.1.1342 15.64 −3.74 0.00137  Bacteria Actino- Actino- Coryne- Coryne- Coryne- bacteria bacteria bacteriales bacteriaceae bacterium 1 New.ReferenceOTU52 1991.15 −2.20 0.01222  Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium sensu stricto 1 HG798451.1.1400 25.03 −2.31 0.00911  Bacteria Firmicutes Bacilli Lacto- Entero- Enterococcus bacillales coccaceae HK557089.3.1395 6043.88 3.13 0.00124  Bacteria Firmicutes Bacilli Lacto- Strepto- Streptococcus bacillales coccaceae GQ448336.1.1418 34.22 3.62 0.02080  Bacteria Firmicutes Negativicutes Seleno- Veillonellaceae Megasphaera monadales KF842598.1.1394 4.25 −4.15 0.02349  Bacteria Bacteroidetes Bacteroidia Bacteroidales Porphyro- Para- monadaceae bacteroides FJ950694.1.1472 1259.08 −3.07 0.00109  Bacteria Proteo- Gamma- Entero- Entero- Escherichia- bacteria proteobacteria bacteriales bacteriaceae Shigella HQ802983.1.1440 40.41 −3.23 0.002803 Bacteria Firmicutes Clostridia Clostridialesa Lachno- Tyzzerella 4 spiraceae GQ448468.1.1366 2058.39 −2.10 0.01036  Bacteria Fusobacteria Fusobacteriia Fuso- Fuso- Fusobacterium bacteriales bacteriaceae JN387556.1.1324 161.95 −3.09 0.01172  Bacteria Firmicutes Clostridia Clostridiales Peptostrepto- Intestinibacter coccaceae

TABLE 5 OTUs with differential abundances between samples day 14 and day 0 in diet responsive dogs (day 14 versus day 0). OTU Base log2Fold- IDs + A2:K39 Mean Change P value Kingdom Phylum Class Order Family Genus Species JRPJ01000002.1034290.1035971 38.04 −3.39 0.034 Bacteria Proteobacteria Epsilonproteobacteria Campylobacterales Helicobacteraceae Helicobacter Ambiguous_taxa New.ReferenceOTU45 32.86 −5.37 0.017 Bacteria Proteobacteria Gammaproteobacteria Aeromonadales Succinivibrionaceae Anaerobiospirillum Ambiguous_taxa GQ006324.1.1342 10.99 −3.41 0.002 Bacteria Actinobacteria Actinobacteria Corynebacteriales Corynebacteriaceae Corynebacterium 1 uncultured bacterium HK555938.1.1357 13.49 5.19 0.034 Bacteria Actinobacteria Coriobacteriia Coriobacteriales Coriobacteriaceae Collinsella uncultured bacterium FJ957551.1.1489 5.64 2.44 0.023 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Sarcina uncultured bacterium FJ957494.1.1454 22.52 2.91 0.001 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium Ambiguous_taxa sensu stricto 1 New.ReferenceOTU52 899.64 −2.89 0.015 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium NA sensu stricto 1 FM865905.1.1392 21.51 −3.54 0.024 Bacteria Firmicutes Clostridia Clostridiales Clostridiaceae 1 Clostridium NA sensu stricto 1 GQ016239.1.1362 12.55 3.10 0.015 Bacteria Firmicutes Erysipelotrichia Erysipelotrichales Erysipelotrichaceae Faecalitalea Ambiguus_taxao HG798451.1.1400 21.63 −1.92 0.023 Bacteria Firmicutes Bacilli Lactobacillales Enterococcaceae Enterococcus Enterococcus durans EU461791.1.1414 5.48 2.98 0.043 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium GU303759.1.1517 22.26 2.50 0.010 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium New.ReferenceOTU114 33.26 3.17 0.002 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium AB506154.1.1541 7.45 2.91 0.008 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium EU774370.1.1398 2.18 3.43 0.029 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium HK557089.3.1395 4123.56 2.62 0.007 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium HQ807346.1.1456 11.8 4.56 2.35E−05 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus uncultured bacterium HQ748204.1.1442 15.13 4.30 4.58E−05 Bacteria Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus unculture bacterium GU179917.1.1382 29.72 2.15 0.044 Bacteria Firmicutes Erysipelotrichia Erysipelotrichales Erysipelotrichaceae Erysipelatoclostridium Ambiguous_taxa GQ448336.1.1418 49.68 4.13 0.014 Bacteria Firmicutes Negativicutes Selenomonadales Veillonellaceae Megasphaera uncultured bacterium DQ804865.1.1390 32.02 3.80 0.030 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae NA NA GQ491757.1.1361 6.02 1.79 0.034 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium New.ReferenceOTU56 33.71 5.05 0.000604 Bacteria Bacteroidetes Bacteroidia Bacteroidales Prevotelaceae Alloprevotella Ambiguous_taxa KF842598.1.1394 4.29 −4.06 0.024 Bacteria Bacteroidetes Bacteroidia Bacteroidales Porphyromonadaceae Parabacteroides NA HQ802052.1.1445 3.40 −3.37 0.010 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides Ambiguous_taxa GX182404.8.1529 2.29 −3.22 0.035 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella NA FJ950694.1.1472 1165.27 −2.81 0.002 Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales Enterobacteriaceae Escherichia-Shigella Escherichia coli GQ448506.1.1374 489.25 2.00 0.011 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium HQ802983.1.1440 25.95 −2.62 0.020 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Tyzzerella 4 NA DQ793824.1.1370 11.84 −3.29 0.009 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae [Ruminococcus] uncultured gauvreauii group bacterium GQ448468.1.1366 2756.51 −2.30 0.013 Bacteria Fusobacteria Fusobacteriia Fusobacteriales Fusobacteriaceae Fusobacterium uncultured bacterium EU774020.1.1361 2.57 −3.07 0.011 Bacteria Fusobacteria Fusobacteriia Fusobacteriales Fusobacteriaceae Fusobacterium uncultured bacterium GQ491183.1.1360 618.96 1.51 0.040 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae NA NA GQ491426.1.1332 506.28 2.55 0.017 Bacteria Firmicutes Clostridia Clostridiales Lachnospiraceae Blautia uncultured bacterium GQ493039.1.1311 82.93 −3.08 0.016 Bacteria Firmicutes Clostridia Clostridiales Peptostreptococcaceae NA NA JN387556.1.1324 135.02 −3.10 0.010 Bacteria Firmicutes Clostridia Clostridiales Peptostreptococcaceae Intestinibacter NA EU775983.1.1288 2.84 2.40 0.008 Bacteria Firmicutes Clostridia Clostridiales Peptostreptococcaceae Peptoclostridium uncultured bacterium

TABLE 6 OTUs with differential abundances between samples of day 0 and day 14 (day 0 versus day 14) for die non-responsive dogs (Fold change >2 and P-value <0.05). Base log2Fold- OTU ID Mean Change P value Kingdom Phylum Class GQ449137.1.1391 461.15 −3.51 0.0187 Bacteria Proteo- Betaproteo- bacteria bacteria HK555938.1.1357 30.11 −6.18 0.0121 Bacteria Actino- Corio- bacteria bacteriia GQ358246.1.1466 302.41 −3.70 0.0182 Bacteria Firmicutes Negativicutes New.ReferenceOTU82 61.40 −3.34 0.0262 Bacteria Firmicutes Clostridia New.ReferenceOTU52 222.71 −4.55 0.0022 Bacteria Firmicutes Clostridia GQ138615.1.1402 321.54 −3.56 0.0059 Bacteria Firmicutes Erysipelo- trichia JN681884.1.1409 384.68 3.09 0.0084 Bacteria Firmicutes Bacilli GU303759.1.1517 48.18 2.96 0.0180 Bacteria Firmicutes Bacilli New.ReferenceOTU114 53.48 4.21 8.04E−05 Bacteria Firmicutes Bacilli EU774881.1.1422 3.84 3.39 0.0242 Bacteria Firmicutes Bacilli AB469559.1.1551 13.56 5.00 0.0016 Bacteria Firmicutes Bacilli HK557089.3.1395 9232.07 4.47 2.88E−05 Bacteria Firmicutes Bacilli EU358719.1.1513 12.02 2.71 0.0180 Bacteria Firmicutes Bacilli HQ748204.1.1442 17.52 2.85 0.0045 Bacteria Firmicutes Bacilli GQ338727.1.1397 9.95 6.38 0.0313 Bacteria Firmicutes Clostridia HQ803964.1.1435 247.41 −2.80 0.0294 Bacteria Firmicutes Clostridia FJ951866.1.1493 7.83 −5.45 0.0177 Bacteria Firmicutes Clostridia EU772870.1.1289 34.63 −4.27 0.0079 Bacteria Fuso- Fusobacteriia bacteria GQ448468.1.1366 4335.90 −4.03 0.0125 Bacteria Fuso- Fusobacteriia bacteria EU774020.1.1361 7.55 −4.76 0.0112 Bacteria Fuso- Fusobacteriia bacteria HQ782658.1.1415 506.92 −6.12 0.0001 Bacteria Fuso- Fusobacteriia bacteria DQ794633.1.1395 23.54 −3.68 0.0209 Bacteria Firmicutes Clostridia FN668375.4306350.4307737 12.13 −5.24 0.0016 Bacteria Firmicutes Clostridia GQ867445.1.1457 24.68 −2.23 0.0150 Bacteria Firmicutes Clostridia OTU ID Order Family Genus Species GQ449137.1.1391 Burkholderiales Alcaligenaceae Sutterella NA HK555938.1.1357 Coriobacteriales Coriobacteriaceae Collinsella uncultured bacterium GQ358246.1.1466 Seleno- Acidamino- Phascolarcto- uncultured monadales coccaceae bacterium Veillonellaceae bacterium New.ReferenceOTU82 Clostridiales Clostridiaceae 1 Clostridium NA sensu stricto 1 New.ReferenceOTU52 Clostridiales Clostridiaceae 1 Clostridium NA sensu stricto 1 GQ138615.1.1402 Erysipelo- Erysipelo- Turicibacter uncultured trichales trichaceae bacterium JN681884.1.1409 Lactobacillales Streptococcaceae Streptococcus NA GU303759.1.1517 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium New.ReferenceOTU114 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium EU774881.1.1422 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium AB469559.1.1551 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium HK557089.3.1395 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium EU358719.1.1513 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium HQ748204.1.1442 Lactobacillales Streptococcaceae Streptococcus uncultured bacterium GQ338727.1.1397 Clostridiales Lachnospiraceae Anaerostipes uncultured bacterium HQ803964.1.1435 Clostridiales Lachnospiraceae Lachno- uncultured clostridium bacterium FJ951866.1.1493 Clostridiales Lachnospiraceae Roseburia NA EU772870.1.1289 Fusobacteriales Fusobacteriaceae Fusobacterium uncultured bacterium GQ448468.1.1366 Fusobacteriales Fusobacteriaceae Fusobacterium uncultured bacterium EU774020.1.1361 Fusobacteriales Fusobacteriaceae Fusobacterium uncultured bacterium HQ782658.1.1415 Fusobacteriales Fusobacteriacea Fusobacterium Ambiguous_taxa DQ794633.1.1395 Clostridiales Lachnospiraceae NA NA FN668375.4306350.4307737 Clostridiales Peptostrepto- NA NA coccaceae GQ867445.1.1457 Clostridiales Lachnospiraceae NA NA

TABLE 7 Comparisons of KEGG pathways between different timepoints in the trial for diet responsive dogs. KEGG pathways Days0vs14_lfc Days0vs14_pvalue Days0vs14_fdr ko00100; Steroid biosynthesis −2.04 0.000209808 0.005895615 ko00312; beta-Lactam resistance 0.44 725E−05 0.005895615 ko00524; Butirosin and neomycin 0.31 0.000164032 0.005895615 biosynthesis ko00630; Glyoxylate and −0.39 0.000125885 0.005895615 dicarboxylate metabolism ko00910; Nitrogen metabolism −0.39 0.000209808 0.005895615 ko03070; Bacterial secretion system −0.46 0.000125885 0.005895615 ko04144; Endocytosis −1.76 0.000209808 0.005895615 ko04912; GnRH signaling pathway −1.76 0.000209808 0.005895615 ko5210, Colorectal cancer −1.63 0.000209808 0.005895615 ko05416; Viral myocarditis −1.63 0.000209808 0.005895615 kko00640; Propanoate metabolism −0.25 0.000267029 0.006821372 ko00010; Glycolysis/Gluconeogenesis 0.19 0.000419617 0.006936017 ko00311; Penicillin and cephalosporin 0.32 0.000419617 0.006936017 biosynthesis ko00920; Sulfur metabolism −0.38 0.000419617 0.006936017 ko04721; Synaptic vesicle cycle −1.15 0.000419617 0.006936017 ko04962; Vasopressin-regulated water −1.15 0.000419617 0.006936017 reabsorption ko05150; Staphylococcus aureus 0.86 0.000335693 0.006936017 infection ko00020; Citrate cycle (TCA cycle) −0.31 0.000644684 0.006967545 ko00052; Galactose metabolism 0.40 0.000644684 0.006967545 ko00240; Pyrimidine metabolism 0.12 0.000522614 0.006967545 ko00410; beta-Alanine metabolism −0.41 0.000644684 0.006967545 ko00473; D-Alanine metabolism 0.29 0.000644684 0.006967545 ko00592; alpha-Linolenic acid −1.38 0.000644684 0.006967545 metabolism ko00633; Nitrotoluene degradation −0.67 0.000644684 0.006967545 ko03410; Base excision repair 0.10 0.000522614 0.006967545 ko04115; p53 signaling pathway −1.57 0.000522614 0.006967545 ko00550; Peptidoglycan biosynthesis 0.18 0.000965118 0.008748331 ko00909; Sesquiterpenoid and −1.52 0.000965118 0.008748331 triterpenoid biosynthesis ko04621; NOD-like receptor 0.43 0.000965118 0.008748331 signaling pathway ko04930; Type II diabetes mellitus 0.17 0.000965118 0.008748331 ko05168; Herpes simplex infection −1.28 0.000965118 0.008748331 ko00281; Geraniol degradation −0.58 0.001411438 0.01166512 ko00540; Lipopolysaccharide −0.53 0.001411438 0.01166512 biosynthesis ko04622; RIG-I-like receptor 0.57 0.001411438 0.01166512 signaling pathway ko00071; Fatty acid metabolism −0.55 0.001693726 0.012863159 ko00120; Primary bile acid 0.51 0.001693726 0.012863159 biosynthesis ko00121; Secondary bile acid 0.51 0.001693726 0.012863159 biosynthesis ko00430; Taurine and hypotaurine −0.32 0.00202179 0.013856655 metabolism ko00590; Arachidonic acid −0.39 0.00202179 0.013856655 metabolism ko05012; Parkinsons disease −0.60 0.00202179 0.013856655 ko05111; Vibrio cholerae pathogenic −0.58 0.00202179 0.013856655 cycle ko00051; Fructose and mannose 0.18 0.002399445 0.014046748 metabolism ko00310; Lysine degradation −0.27 0.002399445 0.014046748 ko00351; DDT degradation −0.89 0.002399445 0.014046748 ko00520; Amino sugar and nucleotide 0.14 0.002399445 0.014046748 sugar metabolism ko00561; Glycerolipid metabolism 0.27 0.002399445 0.014046748 ko01040; Biosynthesis of unsaturated −0.32 0.002399445 0.014046748 fatty acids ko04011; MAPK signaling 0.27 0.002399445 0.014046748 pathway-yeast ko00130; Ubiquinone and other −0.38 0.002838135 0.014241355 terpenoid-quinone biosynthesis ko00380; Tryptophan metabolism −0.58 0.002838135 0.014241355 ko00680; Methane metabolism −0.15 0.002838135 0.014241355 ko02060; Phosphotransferase 0.52 0.002838135 0.014241355 system (PTS) ko04626; Plant-pathogen interaction −0.09 0.002838135 0.014241355 ko04940; Type I diabetes mellitus 0.09 0.002838135 0.014241355 ko05110; Vibrio cholerae infection −1.3 0.002838135 0.014241355 ko05145; Toxoplasmosis −1.25 0.002838135 0.014241355 ko00300; Lysine biosynthesis 0.07 0.003341675 0.015915434 ko02040; Flagellar assembly −0.63 0.003341675 0.015915434 ko04973; Carbohydrate digestion 0.46 0.003341675 0.015915434 and absorption ko05340; Primary immunodeficiency 0.29 0.003917694 0.018347867 ko00511; Other glycan degradation 0.45 0.004577637 0.020098686 ko00791; Atrazine degradation −0.24 0.004577637 0.020098686 ko00983; Drug metabolism-other 0.14 0.004577637 0.020098686 enzymes ko03430; Mismatch repair 0.13 0.004577637 0.020098686 ko00230; Purine metabolism 0.08 0.005329132 0.022689184 ko04260; Cardiac muscle contraction −0.76 0.005329132 0.022689184 ko00620; Pyruvate metabolism −0.06 0.00617981 0.025918306 ko00190; Oxidative phosphorylation −0.11 0.007144928 0.028277814 ko00330; Arginine and proline −0.13 0.007144928 0.028277814 metabolism ko00943; Isoflavonoid biosynthesis 0.55 0.007144928 0.028277814 ko04614; Renin-angiotension system 0.52 0.007144928 0.028277814 ko00062; Fatty acid elongation 0.43 0.008232117 0.030437168 ko02020; Two-component system −0.19 0.008232117 0.030437168 ko03008; Ribosome biogenesis in −0.22 0.008232117 0.030437168 eukaryotes ko04122; Sulfur relay system −0.23 0.008232117 0.030437168 ko04910; Insulin signaling pathway 0.22 0.008232117 0.030437168 ko00471; D-Glutamine and D- 0.15 0.00945282 0.033623321 glutamate metabolism ko00500; Starch and sucrose 0.20 0.00945282 0.033623321 metabolism ko00860; Porphyrin and chlorophyl −0.24 0.00945282 0.033623321 II metabolism ko05132; Salmonella infection −0.33 0.010826111 0.038026714 ko00603; Glycosphingolipid 0.37 0.012359619 0.041844012 biosynthesis-globoseries ko03030; DNA replication 0.08 0.012359619 0.041844012 ko05142; Chagas disease (American −0.63 0.012359619 0.041844012 trypanosomiasis) ko00720; Carbon fixation pathways −0.12 0.014068604 0.046968344 in prokaryotes ko01057; Biosynthesis of type II −0.50 0.014068604 0.045968344 polyketide products ko04146; Peroxisome −0.21 0.014068604 0.045968344

TABLE 8 Comparisons of bile acids between samples at different timepoints for diet responsive dogs. Bile acid Days0vs14_fc Days0vs14_pvalue Days0vs12_fc Days0vs42_pvalue AlphamuricholicAcid 1.74 0.192517572 2.91 0.006835938 DeocycholicAcid 1.74 0.019058892 1.70 0.1015625 GammamuricholicAcid 16.18 0.024390241 18.33 0.014266187 LithocholicAcid 1.50 0.053710938 2.02 0.010826921 OmegamuricholicAcid 8.22 0.022494271 33.55 0.042315275

TABLE 9 Spearman correlations between abundance of OTUs and concentration of Bile acids in diet responsive dogs. Spearman Adjusted Taxa Bile acid correlation Pvalue Pvalue Fusobacterium_uncultured.bacterium Chenodeoxycholic. −0.468045036 0.000533 0.022042405 Acid_primary Bacteroides_NA Chenodeoxycholic. −0.474431034 0.000436 0.022042405 Acid_primary Fusobacterium_uncultured.bacterium Cholic.Acid_primary −0.646842527 3.11E−08 3.86E−06 Fusobacterium_Ambiguous_taxa Cholic.Acid_primary −0.60748964 3.36E−07 2.09E−05 Bacteroides_NA Cholic.Acid_primary −0.592565032 7.64E−07 2.37E−05 Peptoclostridium_uncultured.bacterium Cholic.Acid_primary −0.561738193 3.67E−06 9.11E−05 Megamonas_uncultured.bacterium Cholic.Acid_primary −0.518655946 2.57E−05 0.000532 Bacteroides_uncultured.bacterium Cholic.Acid_primary −0.499276241 5.69E−05 0.001007674 Prevotella.9_uncultured.bacterium Cholic.Acid_primary −0.4938089 7.05E−05 0.001093381 Escherichia.Shigella_Escherichia.coli Cholic.Acid_primary 0.487797189 8.90E−05 0.001226177 Sutterella_NA Cholic.Acid_primary −0.456507609 0.000279 0.003458925 Staphylococcus_Ambiguous_taxa Cholic.Acid_primary 0.418106228 0.000984 0.01108803 Escherichia.Shigella_uncultured.bacterium Cholic.Acid_primary 0.407320492 0.001366 0.014111373 Enterococcus_Enterococcus.durans Cholic.Acid_primary 0.394491205 0.00199 0.018979727 Fusobacterium_uncultured.organism Cholic.Acid_primary −0.391629335 0.00216 0.019129515 Faecalibacterium_uncultured.bacterium Cholic.Acid_primary −0.382982602 0.002755 0.022772609 Clostridium.sensu.stricto.1_NA Cholic.Acid_primary 0.365158178 0.004459 0.030717827 Phascolarctobacterium_uncultured. Cholic.Acid_primary −0.365156122 0.004459 0.030717827 Veillonellaceae.bacterium Erysipelatoclostridium_NA Cholic.Acid_primary 0.369363009 0.003989 0.030717827 Lactobacillus_uncultured.bacterium Cholic.Acid_primary 0.362651494 0.004761 0.030741248 Enterobacter_NA Cholic.Acid_primary 0.361092059 0.004958 0.030741248 Fusobacterium_Ambiguous_taxa Deoxycholic.Acid 0.544270884 5.29E−05 0.00439602 uncultured.bacterium_uncultured. Deoxycholic.Acid −0.536454836 7.09E−05 0.00439602 bacterium Enterococcus_NA Glycochenodeoxycholic. −0.551370353 0.000275 0.034071581 Acid Fusobacterium_uncultured.bacterium Glycocholic.Acid −0.4919575 0.000383 0.047524989 Escherichia.Shigella_Escherichia.coli Glycodeoxycholic.Acid −0.500010873 0.00016 0.006632914 Escherichia.Shigella_uncultured.bacterium Glycodeoxycholic.Acid −0.503678278 0.000141 0.006632914 Escherichia.Shigella_NA Glycodeoxycholic.Acid −0.513698449 9.83E−05 0.006632914 Bacteroides_NA Lithocholic.Acid 0.602216745 1.21E−05 0.001495011 Escherichia.Shigella_Escherichia.coli Lithocholic.Acid −0.505136952 0.000402 0.01214716 Clostridium.sensu.stricto.1_uncultured. Lithocholic.Acid −0.498644383 0.00049 0.01214716 bacterium Alloprevotella_NA Lithocholic.Acid 0.525651352 0.000209 0.01214716 Fusobacterium_uncultured.bacterium Lithocholic.Acid 0.465544021 0.00127 0.022503866 Terrisporobacter_uncultured.bacterium Lithocholic.Acid −0.468902352 0.001158 0.022503866 Fusobacterium_Ambiguous_taxa Taurocholic.Acid −0.597078223 9.46E−07 0.000117322 Bacteroides_uncultured.bacterium Taurocholic.Acid −0.478414977 0.000167 0.006908187 Fusobacterium_uncultured.bacterium Taurocholic.Acid −0.467425093 0.000247 0.007641602 Peptoclostridium_uncultured.bacterium Taurocholic.Acid −0.447367036 0.000485 0.012022892 Prevotella.9_uncultured.bacterium Taurocholic.Acid −0.432173393 0.000788 0.015017592 Catenibacterium_uncultured.bacterium Taurocholic.Acid −0.429812058 0.000848 0.015017592 Bacteroides_NA Taurocholic.Acid −0.419308054 0.001168 0.018102789 Megamonas_uncultured.bacterium Taurocholic.Acid −0.395318261 0.002339 0.032219997 Sutterella_NA Taurocholic.Acid −0.379322634 0.003614 0.044819403 Parasutterella_uncultured.bacterium Taurocholic.Acid 0.372251442 0.004352 0.049062689 Terrisporobacter_uncultured.bacterium Taurolithocholic.Acid −0.47287393 0.000104 0.012896126 Escherichia.Shigella_NA Taurolithocholic.Acid −0.408085979 0.000993 0.046070117 Prevotellacae.UCG.003_uncultured. Taurolithocholic.Acid −0.404397284 0.001115 0.046070117 bacterium

TABLE 10 Comparison of the amount of bile acids in the fecal samples of healthy dogs, diet responsive dogs with CCE and diet non-responsive dogs with CCE. Mean Std. Group Timepoint Bile Acid (nmol/g) CI lower CI upper Error (mg/g) #% DR 0 Deoxycholic 753.4492 385.5109 1121.3876 167.1698 0.295783059 0.029578 DR 14 Deoxycholic 1232.6501 787.5515 1677.7488 209.9616 0.483903915 0.04839 DR 42 Deoxycholic 1229.1665 663.6408 1794.6923 266.7693 0.482536351 0.048254 NDR 0 Deoxycholic 487.2588 −186.5451 1161.0627 242.6858 0.191284162 0.019128 NDR 14 Deoxycholic 736.7662 −124.4111 1597.9436 335.0126 0.289233781 0.028923 NDR 42 Deoxycholic 33.16975 −57.53601 123.87551 32.66975 0.013021515 0.001302 Healthy Deoxycholic 2328.5853 1519.0883 3138.0823 357.8429 0.914137388 0.091414 DR 0 Lithocholic 132.0341 46.31136 217.75684 38.94744 0.049720424 0.004972 DR 14 Lithocholic 209.8332 120.17778 299.48862 42.29218 0.079017434 0.007902 DR 42 Lithocholic 266.74216 153.50886 379.97547 53.41432 0.100447789 0.010045 NDR 0 Lithocholic 56.86471 −39.08019 152.80961 34.55674 0.021413692 0.002141 NDR 14 Lithocholic 74.4107 −46.58682 195.40822 47.07009 0.028021031 0.002802 NDR 42 Lithocholic 1.718967 −1.665428 5.103362 1.218967 0.000647316 6.47E−05 Healthy Lithocholic 601.7535 314.6058 888.9013 126.9354 0.22660388 0.02266 Note: The values <1 nmol/g were under the limit of detection; so an appropriate value 0.5 was used for the calculation.

TABLE 11 16S rRNA Sequences of OTUs in Tables 2-5 OTUs in Table 2 JRPJ01000002.1034290.1035971 AGAGTTTGATCCTGGCTCAGAGTGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGATGAAACTTCTAGCT TGCTAGAAGTGGATTAGTGGCGCACGGGTGAGTAATGCATAGGTAACATGCCCTTTAGTCTGGGATAGCCACTGGA AACGGTGATTAATACTGGATACTCCCTACGGGGGAAAGGGGCTTTCAATAAAGAATTTCTCTTTTTAGTGTTTTGT GTTGTTGGCACAAAATTCTAGTATTTGGAATGAGAAATTGGTGTTGTGAAGCAATTTGTGCGGAGATTAGACTTAG TGTCTGTCGTGTCAGCAAATTGCGAACTCATCGATTTATCATCCAAAGACGAATTTTTTATTGAAAGCCTTCGCTA AAGGATTGGCCTATGTCCTATCAGCTTGTTGGTGAGGTAATGGCTCACCAAGGCTATGACGGGTATCCGGCCTGAG AGGGTGATCGGACACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTAGGGAATATTGCTCAAT GGGGGAAACCCTGAAGCAGCAACGCCGCGTGGAGGATGAAGGTTTTAGGATTGTAAACTCCTTTTGTAAGAGAAGA TTATGACGGTATCTTACGAATAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTT ACTCGGAATCACTGGGCGTAAAGAGCGCGTAGGCGGGTGGTCAAGTCAGATGTGAAATCCTGTAGCTTAACTACAG AACTGCATTTGAAACTGACCATCTAGAGTATGGGAGAGGTAGGTGGAATTCTTGGTGTAGGGGTAAAATCCGTAGA GATCAAGAGGAATACTCATTGCGAAGGCGACCTGCTGGAACATTACTGACGCTGATGCGCGAAAGCGTGGGGAGCA AACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGAATGCTAGTTGTTGTGAGGCTTGTCCTTGCAGTAA TGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATAGACGGGGACCC GCACAAGCGGTGGAGCATGTGGTTTAATTCGATGATACGCGAAGAACCTTACCTAGGCTTGACATTGATAGAATCT ACTAGAGATAGTGGAGTGCCCTTTTAGGGAGCTTGAAAACAGGTGCTGCACGGCTGTCGTCAGCTCGTGTCGTGAG ATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTCCTTAGTTGCTAGCAGTTTGGCTGAGCACTCTAAGGAGA CTGCCTTCGTAAGGAGGAGGAAGGTGAGGACGACGTCAAGTCATCATGGCCCTTACGCCTAGGGCTACACACGTGC TACAATGGGGTGCACAAAGAGATGCAATAGTGTGAGCTGGAGCCAATCTCTAAAACATCTCTCAGTTCGGATTGTA GTCTGCAACTCGACTACATGAAGCTGGAATCGCTAGTAATCGCAAATCAGCAATGTTGCGGTGAATACGTTCCCGG GTCTTGTACTCACCGCCCGTCACACCATGGGAGTTGTATTTGCCTTAAGTCGGAATGCTAAATTGGCTACCGCCCA CGGCAGATGCAGCGACTGGGGTGAAGTCGTAACAAGGTAACCGTAGGTGAACCTGCGGTTG JF920309.1.1340 AGTGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGATGAAGCTTCTAGCTTGCTAGAAGTGGATTAGTGG CGCACGGGTGAGTAAGGTATAGTTAATCTGCCCTACACAAGAGGACAACACCTAGAAATGGGTGCTAATACTCTAT ACTCCTGCTTAACACAAGTTGAGTAGGGAAAGTTTTTCGGTGTAGGATGAGACTATATAGTATCAGCTAGTTGGTA AGGTAAAGGCTTACCAAGGCTATGACGCTTAAGAGGTCTGAGAGGATGATCTCTCACACTGGAACTGAGACACGGT CCAGACTCCTACGGGAGGCAGCAGTAGGGAATATTGCGCAATGGGCGAAAGCCTGACGCAGCAACGCCGCGTGGAG GATGACACTTTTAGGAGCGTAAACTCCTTTTCTTAGGGAAGAATTCTGACGGTACCTAAGGAATAAGCACCGGCTA ACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTACTCGGAATCACTGGGCGTAAAGGGCGCGTAGG CGGATTATCAAGTCTCTTGTGAAATCTAATGGCTTAACCATTAAACTGCTTGGGAAACTGATAGTCTAGAGTGAGG GAGAGGCAGATGGAATTGGTGGTGTAGGGGTAAAATCCGTAGATATCACCAAGAATACCCATTGCGAAGGCGATCT GCTGGAACTCAACTGACGCTAAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCT AAACGATGTATGCTAGTTGTTGGGCTGCTAGTCAGCTCAGTAATGCAGCTAACGCATTAAGCATACCGCCTGGGGA GTACGGTCGCAAGATTAAAACTCAAAGGAATAGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAA GATACGCGAAGAACCTTACCTAGGCTTGATATCCAACAAAGCTTCTAGAGATAGAAGTGTGCTAGCTTGCTAGAAT GTTGAGACAGGTGCTGCACGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACC CACGTATTTAGTTGCTAACACTTCGGGTGAGCACTCTAAATAGACTGCCTTCGTAAGGAGGAGGAAGGTGTGGACG ACGTCAAGTCATCATGGCCCTTATGCCTAGGGCGACACACGTGCTACAATGGCATATACAATGAGACGCAATACCG CGAGGTGGAGCAAATCTATAAAATATGTCCCAGTTCGGATTGTTCTCTGCAACTCGAGAGCATGAAGCCGGAATCG CTAGTAATCGCAAATCAGCCATGTTGCGGTGAATACGTTCCCGGGTCT FJ978526.1.1378 CATGCAAGTCGAACGGTAACATAGAGGAAGCTTGCTTTCTCTGATGACGAGTGGCGGACGGGTGAGTAAGGTCTGG GAAACTGCCTGACAGAGGGGGACAACAACTGGAAACGGTTGCTAATACCGCATACACCCTGAGGGGGAAAGTCGAA AGACGCTGTCAGATGTGCCCAGATGGGATTAGCTAGTAGGTGAGGTAAAGGCTCACCTAGGCGACGATCTCTAGCT GGTCTGAGAGGATGATCAGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATAT TGCACAATGGGGGGAACCCTGATGCAGCCATGCCGCGTGTGTGAAGAAGGCCTTCGGGTTGTAAAGCACTTTCAGA GGGGAGGAAAATGACGTTACCCTCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTG CAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGCAGGCGGTTCTGCAAGTAGGGTGTGAAAGCCCGGGGCTC AACCTCGGAATTGCACTCTAAACTGTGGGACTAGAGTATTGCAGGGGGAGACGGAATTCCAGGTGTAGCGGTGGAA TGCGTAGAGATCTGGAAGAACACCAAAGGCGAAGGCAGTCTCCTGGGCAAATACTGACGCTCATATGCGAAAGCGT GGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTTGATTAGAAGCTTGCTTGTAAGAGTG GGTTTCGCAGCTAACGCGATAAATCAACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGG GGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGACGCAACGCGATGAACCTTACCTGATCTTGACATCGCGAG AATTACTTGTAATGAGTAAGTGCCTTCGGGAACTCGCAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGA GATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCCTTTGTTGCCAGCGGGTAGAGCCGGGAACTCAAAGGA GACTGCCAGTGATAAACTGGAGGAAGGTAGGGATGACGTCAAGTCATCATGGCCCTTACGGTCAGGGCTACACACG TGCTACAATGGGGCGTACAGAGGGAAACGAAACTGCGAGGTGGAGTGGAACCCAGAAAGCGTCCCTAAGTTCGGAT TGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGCAAATCAGAATGTTGCGGTGAATACGTTCC CGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGATTGCACCAGAAGTGGCCAGCCTAACTGCAAAGAGGG CGGTACCACG New.ReferenceOTU45 TACGGAGGGTGCAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGTAGGCGGAAAGGCAAGCAAGATGTGAAA GACCTGGGCTCAACCTGGGTTGGTCATTTTGAACTACCTTTCTAGAGTATTGCAGAGGGAGATGGAATTTCAGGTG TAGCGGTGGAATGCGTAGATATCTGAAAGAACACCAGAGGCGAAGGCGGTCTCCTGGGCAAATACTGACGCTGAGG TGCGAAAGCGTGGGGAGCAAACAGG HK555938.1.1357 ACGGCACCCCTCTCCGGAGGGAAGCGAGTGGCGAACGGCTGAGTAACACGTGGAGAACCTGCCCCCTCCCCCGGGA TAGCCGCCCGAAAGGACGGGTAATACCGGATACCCCCGGGCGCCGCATGGCGCCCGGGCTAAAGCCCCGACGGGAG GGGATGGCTCCGCGGCCCATCAGGTAGACGGCGGGGTGACGGCCCACCGTGCCGACAACGGGTAGCCGGGTTGAGA GACCGACCGGCCAGATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTGCGCAATG GGGGGAACCCTGACGCAGCGACGCCGCGTGCGGGACGGAGGCCTTCGGGTCGTAAACCGCTTTCAGCAGGGAAGAG TCAAGACTGTACCTGCAGAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCGAGCGTT ATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGGCAGGCCGGGGGTCGAAGCGGGGGGCTCAACCCCCC GAAGCCCCCGGAACCTCCGCGGCTTGGGTCCGGTAGGGGAGGGTGGAACACCCGGTGTAGCGGTGGAATGCGCAGA TATCGGGTGGAACACCGGTGGCGAAGGCGGCCCTCTGGGCCGAGACCGACGCTGAGGCGCGAAAGCTGGGGGAGCG AACAGGATTAGATACCCTGGTAGTCCCAGCCGTAAACGATGGACGCTGGGTGTGGGGGGACGATCCCCCCGTGCCG CAGCCNACGCATTAAGCGTCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGC ACAAGCAGCGGAGCATGTGGCTTAATTCGAAGCAACGCGAAGAACCTTACGGCGCATCCCCCCGAGGCCCACGGGG GGTCCGCCGCGTGGGTCAGAGGAGCGCATACGGGAGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGT TAAGTCCCGCAACGAGCGCAACCCCCGCCGCGTGTTGCCATCGGGTGATGCCGGGAACCCACGCGGGACCGCCGCC GTCAAGGCGGAGGAGGGCGGGGACGACGTCAAGTCATCATGCCCCTTATGCCCTGGGCTGCACACGTGCTACAATG GCCGGTACAGAGGGATGCCACCCCGCGAGGGGGAGCGGATCCCGGAAAGCCGGCCCCAGTTCGGATTGGGGGCTGC AACCCGCCCCCATGAAGTCGGAGTTGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATGCGTTCCCGGGCCTTGT ACACACCGCCCGTCACACCACCCGAGTCGTCTGCACCCGAAGTCGCCGGCCCAACCGCAAGGGGG FJ957494.1.1454 TGAGTTTGATCATGGCTCAGGACGAACGCTGGCGGCGTGCCTAACACATGCAAGTCGAGCGATGAAATTTTCTTCG GAAAATGGATTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTATAGAGAGGGATAGCCTTCCGAAAGG GAGATTAATACCTCATAATATCCTAGTATCGCATGATACATGGATTAAAGGAGCAATCCGCTATAGGATGGACCCG CGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCC ACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTG ATGCAGCAACGCCGCGTGAGTGATGACGGTCTTCGGATTGTAAAGCTCTGTCTTTAGGGACGATAATGACGGTACC TAAGGAGGAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTACT GGGCGTAAAGGGAGCGTAGGCGGATCTTTAAGTGGGATGTGAAATACTCGGGCTCAACCTGGGGGCTGCATTCCAA ACTGGGGATCTAGAGTACAGGAGGGGNGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAAC ACCAGTGGCGAAGGCGACTNTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGAT ACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTAGGGGGTGTCAACTCCCCCTGTGCCGCCGCTAACGC ATTAAGTATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGTAGCG GAGCATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCTAGACTTGACATCTTCTGCATTACCCTTAATCGGG GAAGTTCCTTCGGGGACAGAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTC CCGCAACGAGCGCAACCCTTAAGCTTAGTTGCCATCATTAAGTTGGGCACTCTAAGTTGACTGCCGGTGACAAACC GGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGTCTAGGGCTACACACGTGCTACAATGGCAAGTAC AAAGAGAAGCAATACTGTGAAGTGGAGCAAAACTCAAAAACTTGTCTCAGTTCGGATTGTAGGCTGAAACTCGCCT ACATGAAGCTGGAGTTGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGC CCGTCACACCATGAGAGTTGGCAATACCCGAAGTCCGTAAGCTAACCGTAAGGAGGCAGCGGCCGAAGGTAGGGTC AGCGATGGGG New.ReferenceOTU52 TACGTAGGTGGCGAGCGTTATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATGATTAAGTGGGATGTGAAA TACCCGGGCTCAACTTGGGTGCTGCATTCCAAACTGGTTATCTAGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTG TAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGACTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGG DQ797046.1.1403 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCATGCTTAACACATGCAAGTCGAACGGACTGATTCCTTCGG GATGAAAGTTAGTGGCGAACGGGTGAGTAATGTATGAGCAACCTGCCTCTGTCAACGGGATAACAGTTGGAAACGA CTGCTAATACGGTATATGACCACGGCACCGCATGGTGCAGCGGTAAAAGATTTTATCGGACAGAGATGGGCTCATA TCCCATTAGGTAGTTGGTGAGATAACAGCCCACCAAGCCGACGATCAGTAGCCGGTCTGAGAGGATGAACGGCCAC ACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTCCGCAATGGACGAAAGTCTGAC GGAGCAACGCCGCGTGAACGATGAAGGTCTTCGGATTGTAAAGTTCTGTGATCCGGGACGAAGGCATTGATTGAGA ACATTGATTGATGTTGACGGTACCGGAAAAGCAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGG TGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCCGTGCAAGTCCATCTTAAAAGCGTGGGG CTTAACCCCATGAGGGGATGGAAACTGCAGGGCTGGAGTGTCGGAGGGGAAAGTGGAATTCCTAGTGTAGCGGTGA AATGCGTAGAGATTAGGAAGAACACCGGTGGCGAAGGCGACTTTCTAGACGACAACTGACGCTGAGGCGCGAAAGC GTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGGATACTAGGTGTAGGAGGTATCGAC CCCTTCTGTGCCGGAGTTAACGCAATAAGTATCCCGCCTGGGAAGTACGATCGCAAGATTAAAACTCAAAGGAATT GACGGGGGCCCGCACAAGCGGTGGAGTATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCAAGCCTTGACAT TGATCGCAATCCGCAGAAATGCGGAGTTCCTCTTCGGAGGACGAGAAAACAGGTGGTGCACGGCTGTCGTCAGCTC GTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCTTCTGTTGCCAGCACGTAAAGGTGGGAA CTCAGGAGAGACCGCCGCGGACAACGCGGAGGAAGGCGGGGATGACGTCAAGTCATCATGCCCCTTATGGCTTGGG CTACACACGTACTACAATGGGTGCAAACAAAGAGAAGCGAAGTCGCGAGATGGAGCGGACCTCATAAACGCACTCC CAGTTCAGATTGCAGGCTGCAACCCGCCTGCATGAAGTAGGAATCGCTAGTAATCGCGGGTCAGCATACCGCGGTG AATACGTTCCCGGGCCTTGTACACACCGCCCGTCA GQ449092.1.1375 CTGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAAGAGGGTTAGAATGAGAGCTTCGGC AGGATTTCTTTCCATCTTAGTGGCGGACGGGTGAGTAACGTGTGGGCAACCTGCCCTGTACTGGGGGATAATCATT GGAAACGATGACTAATACCGCATGTGGTTCTCGGAAGGCATCTTCTGAGGAAGAAAGGATTTATTCGGTACAGGAT GGGCCCGCATCTGATTAGCTAGTTGGTGAGATAACAGCCCACCAAGGCGACGATCAGTAGCCGACCTGAGAGGGTG ATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGA AAGCCTGATGCAGCAACGCCGCGTGAAGGATGAAGGGTTTCGGCTCGTAAACTTCTATCAATAGGGAAGAAACAAA TGACGGTACCTAAATAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATC CGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGCATGGTAAGCCAGATGTGAAAGCCTTGGGCTTAACCCGAGGAT TGCATTTGGAACTATCAAGCTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATAT TAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGAAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC AGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTCGGGGAGGAATCCTCGGTGCCGTAGC TAACGCAATAAGCACTCCACCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAA GCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAGGCTTGACATCCCGATGACCGTCCTAG AGATAGGACTTCTCTTCGGAGCATCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGT TAAGTCCCGCAACGAGCGCAACCCTTGTCACTAGTTGCTACGAAAGGGCACTCTAGTGAGACTGCCGGTGACAAAC CGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATACAATGGTCGGAA CAGAGGGCAGCGAAGCCGTGAGGCGGAGCCAATCCCAGAAAACCGATCGTAGTCCGGATTGCAGTCTGCAACTCGA CTGCATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACC GCCCGTA AMCI01001631.34.1456 GGCGCACGGGTGAGTAACACGTATCCAACCTGCCGATAACTCGGGGATAGCCTTTCGAAAGAAAGATTAATACCCG ATGGCATGTAAAGACCTCCTGGTCTTTACATTAAAGAATTTCGGTTATCGATGGGGATGCGTTCCATTAGATAGTA GGCGGGGTAACGGCCCACCTAGTCCACGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGACA CGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGACGCGAGTCTGAACCAGCCAAGTAGCGT GAAGGAAGACTGCCCTATGGGTTGTAAACTTCTTTTATACGGGAATAAAGTATTCCACGTGTGGGATTTTGTATGT ACCGTATGAATAAGGATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATCCGAGCGTTATCCGGATTT ATTGGGTTTAAAGGGAGCGTAGGTGGAAGATTAAGTCAGCCTGTGAAAGTTTGCGGCTTAACCGTAAAATTGCAGT TGATACTGGTTTTCTTGAGTGCAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAA GAACTCCGATTGCGAAGGCAGCTCACTGGACTGTAACTGACACTGATGCTCGAAAGTGTGGGTATCAAACAGGATT AGATACCCTGGTAGTCCACACAGTAAACGATGAATACTCGCTGTTTGCGATATACAGTAAGCGGCCAAGCGAAAGC GTTAAGTATTCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGAG GAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGCTTAAATTACACCTGAATAGATTGGAAACAT TTTAGCCGCAAGGCAGGTGTGAAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGCCA TAACGAGCGCAACCCTTATCTTCAGTTACTAACAGTTATAGCTGAGGACTCTGAAGAGACTGCCGTCGTAAGATGT GAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACGTCCGGGGCTACACACGTGTTACAATGGGGGGTACA GAAGGCTGCTACCTGGCGACAGGATGCCAATCCTTAAATCCTCTCTCAGTTCGGACTGGAGTCTGCAACCCGACTC CACGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGC CCGTCAAGCCATGAAAGCCGGGGGTACCTGAAGTGCGTAACCGCAAGGAGCGTCCTAGGGTAAAACTGGTAATTGG GGCTAAGTCGTAACAAGGTAGCCGTACCGGAAGGTGCGGCTG KF842598.1.1394 AGAGTTTGATCCTGGCTCAGGATGAACGCTAGCGACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGATTTGTA GCAATACAGATTGATGGCGACCGGCGCACGGGTGAGTAACGCGTATGCAACTTACCTATCAGAGGGGGATAGCCCG GCGAAAGTCGGATTAATACCCCATAAAACAGGGGTCCCGCATGGGAATATTTGTTAAAGATTCATCGCTGATAGAT AGGCATGCGTTCCATTAGGCAGTTGGCGGGGTAACGGCCCACCAAACCGACGATGGATAGGGGTTCTGAGAGGAAG GTCCCCCACATTGGTACTGAGACACGGACCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGCCGA GAGGCTGAACCAGCCAAGTCGCGTGAAGGAAGAAGGATCTATGGTCTGTAAACTTCTTTTATAGGGGAATAAAGTG GAGGACGTGTCCTTTTTTGTATGTACCCTATGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGG AGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGTGGTGATTTAAGTCAGCGGTGAAAGTTTG TGGCTCAACCATAAAATTGCCGTTGAAACTGGGTTACTTGAGTGTGTTTGAGGTAGGCGGAATGCGTGGTGTAGCG GTGAAATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCTTACTAAACCATAACTGACACTGAAGCACGA AAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCAGTAAACGATGATTACTAGGAGTTTGCGATAC AATGTAAGCTCTACAGCGAAAGCGTTAAGTAATCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATT GACGGGGGCCCGCACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGTTTGAACG TAGTCTGACCGGAATGGAAACACTCCTTCTAGCAATAGCAGATTACAAGGTGCTGCATGGTTGCCTCAACTCCGGC CCGGAAGGTCCGGCTTAATTGCCATAACAAGCGCACCCTTTTACCAAGGTTCAAACAGGTGAAGCTTGAAGACTCT GTGGAACCTCCCCCCTAACCTGTGAGAAGAAGTGGGGATACACTCAATAAACCACGGCCCTTAATCCCGGGGGGAA CACTGGTTACAATGGGTTGGGAAAGGGGGCTTCCTGGCGACAGGATGCTAATCTCCAAACCATGTCTCAGTTCGGA TCGGAGTCTGCAACTCGACTCCGTGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTT CCCGGGCCTTGTACACACCGCCCGTC HQ793763.1.1451 GATGAACGCTAGCTACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGGTCTTAGCTTGCTAAGGCTGATGGCGA CCGGCGCACGGGTGAGTAACACGTATCCAACCTGCCGTCTACTCTTGGCCAGCCTTCTGAAAGGAAGATTAATCCA GGATGGGATCATGAGTTCACATGTCCGCATGATTAAAGGTATTTTCCGGTAGACGATGGGGATGCGTTCCATTAGA TAGTAGGCGGGGTAACGGCCCACCTAGTCAACGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTG AGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGGCGCGAGCCTGAACCAGCCAAGT AGCGTGAAGGATGACTGCCCTATGGGTTGTAAACTTCTTTTGTCCGGGAATAAAACCGCCTACGTGTAGGCGCTTG TATGTACCGGTACGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATGCGAGCGTTATCC GGATTTATTGGGTTTAAAGGGAGCGCAGACGGGTTTTTAAGTCAGCTGTGAAAGTTTGGGGCTCAACCTTAAAATT GCAGTTGATACTGGAGACCTTGAGTGCAGTTGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATC ACGAAGAACTCCGATTGCGAAGGTAGCTTGCTAAAGTGTAACTGACGTTCATGCTCGAAAGTGTGGGTATCAAACA GGATTAGATACCCTGGTAGTCCACACGGTAAACGATGGATACTCGCTGTTGGCGATATACGGTCAGCGGCTTAGCG AAAGCGTTAAGTATCCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAG CGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGCTTAAATTGCACTGGACTATTCTGGA AACAGGATATTCTTCGGACCAGTGTGAAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAG TGCCATAACGAGCGCAACCCTTGCTGCCAGTTACTAACAGGTAATGCTGAGGACTCTGGCGGGACTGCCATCGTAA GATGCGAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACGTCCGGAGCTACACACGTGTTACAATGGTAG GTACAGAGGGTAGCTACCCAGCGATGGGATGCGAATCTCGAAAGCCTATCTCAGTTCGGATTGGAGGCTGAAACCC GCCTCCATGAAGTTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCCGGGCCTTGTACAC ACCGCCCGTCAAGCCATGGGAGCCGGGGGTACCTGAAGTACGTAACCGCAAGGATCGTCCTAGGGTAAAACTGGTG ACTGGGG DQ113765.1.1450 GATGAACGCTAGCTACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGAAGTTTGCTTGCAAACTTTGATGGCGA CCGGCGCACGGGTGAGTAACGCGTATCCAACCTCCCGCATACTCGGGGATAGCCTTCTGAAAGGAAGATTAATACC CGATGGTATCTTAAGCGCACATGCAATTAAGATTAAAGAATTTCGGTATGCGATGGGGATGCGTTCCATTAGGTAG TAGGCGGGGTAACGGCCCACCTAGCCATCGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGA CACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGGCGCGAGCCTGAACCAGCCAAGTAGC GTGAAGGATGACTGCCCTATGGGTTGTAAACTTCTTTTGTCCGGGAATAAAACCGCCTACGTGTAGGCGCTTGTAT GTACCGGTACGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATGCGAGCGTTATCCGGA TTTATTGGGTTTAAAGGGAGCGCAGACGGGTTTTTAAGTCAGCTGTGAAAGTTTGGGGCTCAACCTTAAAATTGCA GTTGATACTGGAGACCTTGAGTGCAGTTGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACG AAGAACTCCGATTGCGAAGGCAGCTTGCTAAAGTGTAACTGACGTTCATGCTCGAAAGTGTGGGTATCAAACAGGA TTAGATACCCTGGTAGTCCACACGGTAAACGATGGATACTCGCTGTTGGCGATATACGGTCAGCGGCTTAGCGAAA GCGTTAAGTATCCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGG AGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGCTTAAATTGCACTGGACTTTCCCGGAAAC GGGATTTTCTTCGGACCAGTGTGAAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGC CATAACGAGCGCAACCCTTGCTGCCAGTTACTAACAGGTAATGCTGAGGACTCTGGCGGGACTGCCATCGTAAGAT GCGAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACGTCCGGGGCTACACACGTGTTACAATGGGGGGTA CAGAAGGCCGCTACCCGGCAACGGGATGCCAATCTCCAAAACCCCTCTCAGTTCGGACTGGAGTCTGCAACCCGAC TCCACGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCACGGCGCGGTGAATACGTTCCCGGGCCTTGTACACACC GCCCGTCAAGCCATGAAAGCCGGGGGTACCTGAAGTGCGTAACCGCAAGGAGCGCCCTAGGGTAAAACTGGTAATT GGGGCT ACBW01000012.3536.5054 AGAGTTTGATCCTGGCTCAGGATGAACGCTAGCTACAGGCTTAACACATGCAAGTCGAGGGGCAGCGGGATTGAAG CTTGCTTCAATTGCCGGCGACCGGCGCACGGGTGAGTAACGCGTATCCAACCTTCCGCTTACTCGGGGATAGCCTT TCGAAAGAAAGATTAATACCCGATGGTATCTTAAGCACGCATGAGATTAAGATTAAAGATTTATCGGTAAGCGATG GGGATGCGTTCCATTAGGCAGTTGGCGGGGTAACGGCCCACCAAACCTACGATGGATAGGGGTTCTGAGAGGAAGG TCCCCCACATTGGAACTGAGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGGCGAG AGCCTGAACCAGCCAAGTAGCGTGAAGGATGACGGCCCTACGGGTTGTAAACTTCTTTTGTGCGGGAATAAAGGAA CCTACGTGTAGGTTTTTGCATGTACCGTAACGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGG AGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGACGGGTTTTTAAGTCAGCTGTGAAAGTTTG GGGCTCAACCTTAAAATTGCAGTTGAAACTGGAGACCTTGAGTACGGTTGAGGCAGGCGGAATTCGTGGTGTAGCG GTGAAATGCTTAGATATCACGAAGAACCCCGATTGCGAAGGCAGCCTGCTAAGCCGCCACTGACGTTGAGGCTCGA AAGTGCGGGTATCAAACAGGATTAGATACCCTGGTAGTCCGCACGGTAAACGATGGATACTCGCTGTTGGCGATAG ACAGTCAGCGGCCAAGCGAAAGCGTTAAGTATCCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATT GACGGGGGCCCGCACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGCTTGAACT GCAGTGGAATTATCCGGAAACGGATAAGCGAGCAATCGCCGCTGTGGAGGTGCTGCATGGTTGTCGTCAGCTCGTG CCGTGAGGTGTCGGCTTAAGTGCCATAACGAGCGCAACCCTTGCTGCCAGTTACTAACAGGTCATGCTGAGGACTC TGGCAGGACTGCCATCGTAAGATGCGAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACGTCCGGGGCTA CACACGTGTTACAATGGGGAGTACAGAGGGCAGCTACCGGGCGACCGGATGCGAATCCCGAAAGCTCCTCTCAGTT CGGACTGGAGTCTGCAACCCGACTCCACGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCACGGCGCGGTGAATA CGTTCCCGGGCCTTGTACACACCGCCCGTCAAGCCATGAAAGCCGGGGGTACCTGAAGTACGTAACCGCGAGGATC GTCCTAGGGTAAAACCGGTAATTGGGGCTAAGTCGTAACAAGGTAGCCGTACCGGAAGGTGCGGCTG HK693629.1.1491 AGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAGAAACATTTTAATG AAGCTTCGGCAGATTTAGTTTGTTTCTAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCACACTGGGGG ATAACAGTCAGAAATGACTGCTAATACCGCATAAGCGCACGGAACCGCATGGTTTTGTGTGAAAAACTCCGGTGGT GTGAGATGGACCCGCGTTGGATTAGCCAGTTGGCAGGGTAACGGCCTACCAAAGCGACGATCCATAGCCGGCCTGA GAGGGTGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAA TGGGGGAAACCCTGATGCAGCGACGCCGCGTGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAG ATAATGACGGTACCTGACTAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGT TATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGAGCAGCAAGTCTGATGTGAAAGGCAGGGGCTCAACCCCT GGACTGCATTGGAAACTGTTGATCTTGAGTACCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAG ATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGC AAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTCGGGTGGCAGAGCCATTCGGTG CCGCAGCAAACGCAGTAAGTATTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACC CGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCCTCTGACC GGTCCTTAACCGGACCTTTCCTTCGGGACAGAGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGAT GTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCCCCAGTAGCCAGCATTTAAGGTGGGCACTCTGAGGAGACT GCCAGGGATAACCTGGAGGAAGGCGGGGATGACGTCAAATCATCATGCCCCTTATGATTTGGGCTACACACGTGCT ACAATGGCGTAAACAAAGGGAAGCAGAGCGGTGACGCCGAGCAAATCCCAAAAATAACGTCCCAGTTCGGACTGCA GTCTGCAACTCGACTGCACGAAGCTGGAATCGCTAGTAATCGCGGATCAGAATGCCGCGGTGAATACGTTCCCGGG TCTTGTACACACCGCCCGTCACACCATGGGAGTCAGTAACGCCCGAAGTCAGTGACCTAACCGAAAGGGAGGAGCT GCCGAAGGCGGGACGGATGACTGGGGTGAAGTCGTAACAAGGTAACC JQ208053.1.1336 GATGAACGCTGACAGAATGCTTAACACATGCAAGTCTACTTGAATTCACTTCGGTGATAGTAAGGTGGCGGACGGG TGAGTAACACGTAAAGAACTTGCCTTACAGTCTGGGACAACTATTGGAAACGATAGCTAATACCGGATATTATGCG AGAGTCGCATGACTCTTGTATGAAAGCTATATGCGCTGTAAGAGAGCTTTGCGTCCCATTAGCTAGTTGGTGAGGT AACGGCTCACCAAGGCCACGATGGGTAGCCGGCCTGAGAGGGTGAACGGCCACAAGGGGACTGAGACACGGCCCTT ACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGACCAAAAGTCTGATCCAGCAATTCTGTGTGCACGAT GACGGTCTTAGGATTGTAAAGTGCTTTCAATTGGGAAGAAAAAAATGACGGTACCAATAGAAGAAGCGACGGCTAA ATACGTGCCAGCAGCCGCGGTAATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGCGTCTAGGT GGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCTAACTAGAGTATCGGA GAGGTGGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATAGAGAAGTCAGCTCAC TGGACGAATACTGACACTGAAGCGCGAAAGCATGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCATGCTGTAA ACGATGATTACTAAGCGTCGGGGGTCGAACCTCGGCACTCAAGCTAACGCGATAAGTAATCCGCCTGGGGAGTACG TACGCAAGTATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGTGGTGGAGCATGTGGTTTAATTCGACGCAAC GCGAGGAACCTTACCAGCGTTTGACATCCTAGGAATGAGAAAGAGATTTCTTAGTGCTCCTTCGGGAGAACCTAGA GACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTAT TGTATGTTGCCATCATTAAGTTGGGCACTCATGCGATACTGCCTGCGATGAGCAGGAGGAAGGTGGGGATGACGTC AAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTACAATGGGCAGTACAGAGAGAAGCAAATCTGCGAGG AGGAGCAAATCTCACAAAGCTGTTCGTAGTTCGGATTGTACTCTGCAACTCGAGTACATGAAGTTGGAATCACTAG TAATCGCAAATCAGCTATGTTGCGGTGAATACGTTCTCGGGTCT GQ493166.1.1359 GATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGGAAACATTTTAATGAAGCTTCGGCAGATTTAGCT TGTTTCTAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCACACTGGGGGATAACAGTCAGAAATGACTG CTAATACCGCATAAGCGCACGGAACCGCATGGTTTTGTGTGAAAAACTCCGGTGGTGTGAGATGGACCCGCGTTGG ATTAGCCAGTTGGCAGGGTAACGGCCTACCAAAGCGACGATCCATAGCCGGCCTGAGAGGGTGAACGGCCACATTG GGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAG CGACGCCGCGTGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGACGGTACCTGACTA AGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACTGGGTGT AAAGGGAGCGTAGACGGAATGGCAAGTCTGATGTGAAAGGCAGGGGCTCAACCCCTGGACTGCATTGGAAACTGTC AGTCTTGAGTACCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGT GGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTG GTAGTCCACGCCGTAAACGATGAATACGAGGTGTCGGGTGGGCAAAGCCATTCGGTGCCGCAGCAAACGCAAAAAG TAATCCCACCTGGGGGAGTACGTTCCCAAGAATGAAACTCAAAGGAAATAGCGGGGACCCGCACAAGCGGTGGAGC ATGTGGTGTATTTGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCCTCTGACCGGTCCTTAACCGGACCTC TCCTTCGGGACAGGGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA ACGAGCGCAACCCCTATCCTTAGTAGCCAGCATCTGAGGTGGGCACTCTGAGGAGACTGCCAGGGATAACCTGGAG GAAGGCGGGGAGGACGTCAAATCATCATGCCCCCTATGATTTGGGCTACACACGTGCTACAATGGCGTAAACAAAG GGAAGCAGAGCGGTGACGCCGAGCAAATCCCAAAAATAACGTCCCAGTTCGGACTGCAGTCTGCAACTCGACTGCA CGAAGCTGGAATCGCTAGTAATCGCGGATCAGAATGCCGCGGTGAATAAAAGCCCGGGTCTTGCACT GQ448486.1.1387 AGAGTTTGATCATGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGGAATTACTTTATTG AAGCTTTGGTCGATTTAATTTAATTATAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTTATACAGGGGG ATAACAGTCAGAAATGGCTGCTAATACCGCATAAGCGCACAGAGCTGCATGGCTCAGTGTGAAAAACTCCGGTGGT ATAAGATGGACCCGCGTTGGATTAGTTGGTTGGTGGGGTAACGGCCCACCAAGGCGACGATCCATAGCCGGCCTGA GAGGGTGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCATACGGGAGGCAGCAGTGGGGAATATTGCACAA TGGGGGAAACCCTGATGCAGCGACGCCGCGTGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAG ATAGTGACGGTACCTGACTAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGT TATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTGTGGCAAGTCTGATGTGAAAGGCATGGGCTCAACCTGT GGACTGCATTGGAAACTGTCATACTTGAGTGCCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAG ATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGC AAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTCGGGTGGCAAAGCCATTCGGTG CCGTCGCAAACGCAGTAAGTATTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACC CGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCGCCTGACC GATCCTTAACCGGATCTTTCCTTCGGGACAGGCGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGAT GTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCCTCAGTAGCCAGCATTAAGTTGGGCACTCATGCGATACTG CCTGCGATGAGCAGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTA CAATGGGTAGTACAGAGAGTCGCAAACCTGCGAGGGGGAGCTAATCTCAGAAAACTATTCTCAGTTCGGATTGTAC TCTGCAACTCGAGTACATGAAGTTGGAATCGCTAGTAATCGCAAATCAGCTATGTTGCGGTGAATACGTTCTCGGG TCTTGCACTCACCGCCCGT GQ491426.1.1332 GCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAGCGAAGCACTTGCCATTGACTCTTCGGAAGAT TTGGCATTTGACTGAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACGGGGGAATAACAGTTAGAA ATGGCTGCTAATGCCGCATAACCGCACAGGACCGCATGGACTGGTGTGAAAAACTGAGGTGGTATGAGATGGGCCC GCGTCTGATTAGGTTAGTTGGCGGGGTAACGGCCCACCAAGCCGACGATCAGTAGCCGACCTGAGAGGGACCGGCC ACATTGGGACTGAGACATGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGAGGAAACTCTG ATGCAGCGACGCCGCATGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGACGGTACC TGACTAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACT GGGTGTAAAGGGAGCGTAGACGGACGGGCAAGTCTGATGTGAAAGCCCGGGGCTTAACCCCGGGACTGCATTGGAA ACTGTCCATCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAAC ACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGAT ACCCTGGTAGTCCACGCCGTAAACGATCAATAATGGGTGTCGGGTTGCAAAGCAATCCGGTGCCGCAGCAAACGCA GTAAGTATTCCCCCTCGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAAGGGACGGGGATCCGCACAAGCGGCGG AGCATGTGGTTTAATTAGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCTGCCTGACCGTTCCTTAACCGGA ACTATCTTTCGGGACAGGCAAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCCTGTCCTTAGTAGCCAGCAGTCCGGCTGGGCACTCTAGGGAGACTGCCGGGGGTAACCC GGAGGAAGGCGGGGAGGAGGTCAAATCATCATGCCCCCCCTGATTTGGGCTACACACGTGGTACAATGGCGTAAAC AAAGGGAAGCGGAGTGGTGACGCTGAGCAAATCTCAAAAATAACGTCCCACTTCGGACTGCAGTCTGCAACTCGAC TGCACGAAGCTGGAATCGCTAGTAATCGCGAATCAGAATG New.ReferenceOTU54 TACGTAGGGGGCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGTCTGATGTGAAA GGCAGGGGCTCAACTCCTGGACTGCATTGGAAACTGCCAGGCTTGAGTGCCGGAGGGGTAAGCGGAATTCCTAGTG TAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGG CTCGAAAGCGTGGGGAGCAAACAGG JN387556.1.1324 CGTAAGTAACCTGCCCTGTACACACGGATAACATACCGAAAGGTATGCTAATACGGGATAATATATTTTGATCGCA TGGTCGAGATATCAAAGCTCCGGCGGTACACCAGGGACCCCCGACAGAGGAGCTAGTTGGTAGTAATGTCACCAAG GCGACGATCAGAAGCCGAACTGAGAGGGGGATCCGCACATGACTGAGACACGGTCAAACTCCTACGGGAGGCAGCA GTGGGGAATATGCCAATGGGCGAAAGCTGATGCAGCACGCGCGTGAGCGATGAGGCTCGGGTCGTAAAGCTCGTCT CAAGGAAGATAATGACGGTACTTGAGGAGGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGG GCTAGCGTTATCCGGAATTACTGGGCGTAAAGGGTGCGTAGGCGGTCTTTCAAGTCAGGAGTGAAAGGCTACGGCT CAACCGTAGTAAGCTCTTGAAACTGTAAGACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAA TGCGTAGATATTAGGAGGAACACCAGTTGCGAAGGCGGCTCTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGT GGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTACTAGCTGTCGGAGGTTACCCCCT TCGGTGGCGCAGCTAACGCATTAAGTACTCCGCCTGGGAAGTACGCTCGCAAGAGTGAAACTCAAAGGAATTGACG GGGACCCGCACAAGTAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTAAGCTTGACATCCCA CTGACCCTTCCCTAATCGGAAGCTTCCCTTCGGGACAGTGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTC GTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGCCTTTAGTTGCCAGCATTAAGTTGGGCACTCTAGA GGGACTGCCAGGGATAACCCGGAGGAGTGGGGATGACGTCAAATCATCATGCCCTTATGCTAGGCTACACACGTGC TACAATGGGTGGTCAGAGGCCAGCCAGTCGTGAGGCCGAGCTATCCCATAAGCCATTCTCGTCCGGATTGTAGGCT GAACTCGCCTACATGAGCTGGAATTACAAGTATGCGATCGATGCTGCGTGATGCGTCCGGGTCTTGTACACACCGC CCGTCACACCATGGGAGTTGGGGGCGCCCGAAGCCGGATTGCTAACCTTTTGGAAGCGTCCGTCGAAGGTGAAACC AATAACTGGGGTGAAGTCGTAACAAGGTAACC OTUs in Table 3 GQ006324.1.1342 GACGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGAAAGGCCCCAGCTCGCTGGGGTACTCGAGTGGCG AACGGGTGAGTAACACGTGGGTGATCTGCCTTGCACTCTGGGATAAGCTTGGGAAACTGGGTCTAATACCGGATAT GAACTGCCTTTAGTGTGGTGGTTGGAAAGTTTTTTCGGTGCAAGATGAGCTCGCGGCCTATCAGCTTGTTGGTGGG GTAATGGCCTACCAAGGCGTCGACGGGTAGCCGGCCTGAGAGGGTGTACGGCCACATTGGGACTGAGATACGGCCC AGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGGAAGCCTGATGCAGCGACGCCGCGTGGGGGA TGACGGCCTTCGGGTTGTAAACTCCTTTCGACAGGGACGAAGCTTTTTGTGACGGTACCTGTATAAGAAGCACCGG CTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTGTCCGGAATTACTGGGCGTAAAGAGCTCGT AGGTGGTTTGTCGCGTCGTCTGTGAAATTCCGGGGCTTAACTCCGGGCGTGCAGGCGATACGGGCATAACTTGAGT ACTGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCG GGTCTCTGGGCAGTAACTGACGCTGAGGAGCGAAAGCATGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCATG CCGTAAACGGTGGGCGCTAGGTGTGGGTTTCCTTCCACGGGATCCGTGCCGTAGCTAACGCATTAAGCGCCCCGCC TGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGTGGATTAA TTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATACACTGGATCGGGCTAGAGATAGTCTTTCCCTTTGTGG CTGGTGTACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAAC CCTTGTCTTATGTTGCCAGCATTTGGTTGGGGACTCATGAGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGAT GACGTCAAATCATCATGCCCCTTATGTCCAGGGCTTCACACATGCTACAATGGTCGGTACAACGCGCAGCGACACT GTGAGGTGGAGCGAATCGCTGAAAGCCGGCCTTAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGT CGCTAGTAATCGCAGATCAGCAATGCTGCGGTGAATACGTTCCCGGGCCT New.ReferenceOTU52 TACGTAGGTGGCGAGCGTTATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATGATTAAGTGGGATGTGAAA TACCCGGGCTCAACTTGGGTGCTGCATTCCAAACTGGTTATCTAGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTG TAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGACTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGG HG798451.1.1400 CTTGTGTCACCAACCATAGGGAGGGGGAAAACATGGAAACGGGGTTCATACCGCATAACTTTTTTAGCCCAATGCA TAAGAAGAAAGGCCTTTCGGGTTTCGGTAAAGGAGGCCCCCGCGGCTCTTATAGTGTGTGTGGAAGTAACCGCTTC CACAAGGCCCAGGTTTCATACCCGACTGGAGAGTGTGTTCGCCACACTGGGGAAAGGACCCCCGGCCCAGTCTCTC TAGGGGAGGCAGCAGTAGGAATTTTCGGCAAAGGAAAAAATTTCTGACCGAACAACGCCGGTTGAATGAAGAAGTT TTTCGGATCGAAAAACTCTGTTGTTAGAGAAGAACAAGGACGTTAGTAACTGAACGTCCCCTGACGGTATCTAACC AGAAAGCCACGGCTAATTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCG TAAAGCGAGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGG GAGACTTGAGTGCAGAAGAGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCAG TGGCGAAGGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCT GGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCAAACGCATTAA GCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCA TGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTTTGACCACTCTAGAGATAGAGCTTT CCCTTCGGGGACAAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA ACGAGCGCAACCCTTATTGTTAGTTGCCATCATTTAGTTGGGCACTCTAGCGAGACTGCCGGTGACAAACCGGAGG AAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGAAGTACAACGA GTCGCTAGACCGCGAGGTCATGCAAATCTCTTAAAGCTTCTCTCAGTTCGGATTGCAGGCTGCAACTCGCCTGCAT GAAGCCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGT CACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTTGGAGCCAGCCGCCTAAGGTGGGATAGAT GATTGGGGTGAAGTCGTAACCAACGTATGCC HK557089.3.1395 AGACTTTAGCTTGCTAAAGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGA TAACTATTGGAAACGATAGCTAATACCGCATAACAGCATTTAACCCATGTTAGATGCTTGAAAGGAGCAATTGCTT CACTAGTAGATGGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGAC CTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCG GCAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGA GAAGAACGTGTGTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAG TCTGAAGTTAAAGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGG AATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAAC TGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGC TAGGTGTTAGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAA GGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGA ACCTTACCAGGTCTTGACATCCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACTGTGAGACTTGAGGGCA GAAGGGTAGAGTGCACTTGTATGGGGAGCTGTGGAATGCGTTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCC ATCATTAAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATG CCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTACAACGAGTCGCGAGTCGGTGACGGCAAGCAAATC TCTTAAAGCCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGAT CAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAA GTCGGTGAGGTANCCTTTTAGGAGC GQ448336.1.1418 AGAGTTTGATCATGGCTCAGGACGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAGAAGAGATGAGAAG CTTGCTTCTTATCTCTTCGAGTGGCAAACGGGTGAGTAACGCGTAAGCAACCTGCCCTTCAGATGGGGACAACAGC TGGAAACGGCTGCTAATACCGAATACGTTCTTTTTGTCGCATGGCAGAGGGAAGAAAGGGAGGCTCTTCGGAGCTT TCGCTGAAGGAGGGGCTTGCGTCTGATTAGCTAGTTGGAGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCCGG TCTGAGAGGATGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTC CGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAACGATGACGGCCTTCGGGTTGTAAAGTTCTGTTATACG GGACGAATGGCGTAGCGGTCAATACCCGTTACGAGTGACGGTACCGTAAGAGAAAGCCACGGCTAACTACGTGCCA GCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCGCGCAGGCGGCGTCGTAA GTCGGTCTTAAAAGTGCGGGGCTTAACCCCGTGAGGGGACCGAAACTGCGATGCTAGAGTATCGGAGAGGAAAGCG GAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAAGCGGCTTTCTGGACGACAA CTGACGCTGAGGCGCGAAAGCCAGGGGAGCAAACGGGATTAGATACCCCGGTAGTCCTGGCCGTAAACGATGGATA CTAGGTGTAGGAGGTATCGACCCCTTCTGTGCCGGAGTTAACGCAATAAGTATCCCGCCTGGGGAGTACGGCCGCA AGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAACATGTGGTTTAATTCGATGATACGCGAGG AACCTTACCCGGGCTTAAATTGCAGTGGAATGATGTGGAAACATGTCAGTGAGCAATCACCGCTGTGAAGGTGCTG CATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGCCATAACGAGCGCAACCCTTATCTTCAGTTACT AACAGGTCATGCTGAGGACTCTGGAGAGACTGCCGTCGTAAGATGTGAGGAAGGTGGGGATGACGTCAAATCAGCA CGGCCCTTACGTCCGGGGCTACACACGTGTTACAATGGGGGGTACAGAGGGCCGCTACCACGCGAGTGGATGCCAA TCCCAAAAACCTCTCTCAGTTCGGACTGGAGTCTGCAACCCGACTCCACGAAGCTGGATTCGCTAGTAATCGCGCA TCAGCCACGGCGCGGTGAATACGTTCCCGGGCCTTGCACTCACCGCCCGT KF842598.1.1394 AGAGTTTGATCCTGGCTCAGGATGAACGCTAGCGACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGATTTGTA GCAATACAGATTGATGGCGACCGGCGCACGGGTGAGTAACGCGTATGCAACTTACCTATCAGAGGGGGATAGCCCG GCGAAAGTCGGATTAATACCCCATAAAACAGGGGTCCCGCATGGGAATATTTGTTAAAGATTCATCGCTGATAGAT AGGCATGCGTTCCATTAGGCAGTTGGCGGGGTAACGGCCCACCAAACCGACGATGGATAGGGGTTCTGAGAGGAAG GTCCCCCACATTGGTACTGAGACACGGACCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGCCGA GAGGCTGAACCAGCCAAGTCGCGTGAAGGAAGAAGGATCTATGGTCTGTAAACTTCTTTTATAGGGGAATAAAGTG GAGGACGTGTCCTTTTTTGTATGTACCCTATGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGG AGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGTGGTGATTTAAGTCAGCGGTGAAAGTTTG TGGCTCAACCATAAAATTGCCGTTGAAACTGGGTTACTTGAGTGTGTTTGAGGTAGGCGGAATGCGTGGTGTAGCG GTGAAATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCTTACTAAACCATAACTGACACTGAAGCACGA AAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCAGTAAACGATGATTACTAGGAGTTTGCGATAC AATGTAAGCTCTACAGCGAAAGCGTTAAGTAATCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATT GACGGGGGCCCGCACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGTTTGAACG TAGTCTGACCGGAATGGAAACACTCCTTCTAGCAATAGCAGATTACAAGGTGCTGCATGGTTGCCTCAACTCCGGC CCGGAAGGTCCGGCTTAATTGCCATAACAAGCGCACCCTTTTACCAAGGTTCAAACAGGTGAAGCTTGAAGACTCT GTGGAACCTCCCCCCTAACCTGTGAGAAGAAGTGGGGATACACTCAATAAACCACGGCCCTTAATCCCGGGGGGAA CACTGGTTACAATGGGTTGGGAAAGGGGGCTTCCTGGCGACAGGATGCTAATCTCCAAACCATGTCTCAGTTCGGA TCGGAGTCTGCAACTCGACTCCGTGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTT CCCGGGCCTTGTACACACCGCCCGTC FJ950694.1.1472 CGCCCTGATTGACGGCTATACACATGCAAGTCGAACGGTAACAGGAAACAGCTTGCTTCTTTGCTGACGAGTGGCG GACGGGTGAGTAATGTCTGGGAAACTGCCTGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAAC GTCGCAAGACCAAAGAGGGGGACCTTCGGGCCTCTTGCCATCGGATGTGCCCAGATGGGATTAGCTAGTAGGTGGG GTAACGGCTCCATCCCTAGGCGAGCCGAATCCTTAGCCTGGTCTGAGAGGAATGACCAGCCACACTGGGACTGAGA ACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCG CGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGAGTAAAGTTAATACCCTTTGCTC ATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTA ATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTTGTTAAGTCAGATGTGAAATCCCCGGGCTCAACCTGGGA ACTGCATCTGATACTGGCAAGCTTGAGTCTCGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAG ATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACGAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAA ACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTGGCTTCC GGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCG CACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGTCTTGACATCCACGGGAAGTT TTCAGAGATGAGAATGTGCCTTCGGGAACCGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATG TTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCAGCGGTCCGGCCGGGAACTCAAAGGAGACTG CCAGTGATAAACTGGAGGAAGGTGGGGATGACGTCCAGGTCATCATGGCCCTTACGAACCAGGGCTACACACGTGC CTACAATGGACGCATCCAAAGAGAGAGCGAACCCTGCCCGCGAGAGCAAGCGGACCTCATAAAGTGCGTCGTAGTC CGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTGGATCAGAATGCCACGGTGAATAC GTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCAAAAGAAGTAGGTAGCTTAACCTTCGG GAGGGCGCTTACCACTTTGGATGCGAGG HQ802983.1.1440 TAAGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCCTATGAAGCGCTTAAACGGATTTCTTCGGATTGAAGT TTTTGTGACTGAGTGGCGGACGGGTGAGTAACGCGTGGGTAACTTGCCTCATACAGGGGGATAACAGTTAGAAATG ACTGCTAATACCGCATAAGCGCACAGTGCTGCATGGCACAGTGTGAAAAACTCCGGTGGTATGAGATGGACCCGCG TCTGATTAGCTAGTTGGTGGGGTAACGGCCTACCAAGGCGACGATCAGTAGCCGGCCTGAGAGGGTGAACGGCCAC ATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGAT GCAGCGACGCCGCGTGAGCGAAGAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGGAAGAAAATGACGGTACCTG ACTAAGAAGCACCGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGGTGCAAGCGTTATCCGGATTTACTGG GTGTAAAGGGAGCGTAGACGGTTGTGTAAGTCTGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTGGAAAC TATGTAACTAGAGTGTCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACAC CAGTGGCGAAGGCGGCTTACTGGACGATCACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATAC CCTGGTAGTCCACGCCGTAAACGATGACTACTAGGTGTCGGGGCCCATAAGGGCTTCGGTGCCGCAGCAAACGCAA TAAGTATTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGA GCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGGTCTTGACATCCCACTGACCGGACAGTAATGTGTC CTTTCCTCCGGGACAGTGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCC GCAACGAGCGCAACCCCTATCCTTAGTAGCCAGCAGTAAGATGGGCACTCTAGGGAGACTGCCAGGGATAACCTGG AGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACTTGGGCTACACACGTGCTACAATGGCGTAAACAA AGTGAAGCGAAGTCGTGAGGCCAAGCAAATCACAAAAATAACGTCTCAGTTCGGATTGTAGTCTGCAACTCGACTA CAAGAAGCTGGAATCGCTAGTAATCGCAGATCAGAATGCTGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCC CGTCACACCATGGGAGTCGAAAATGCCCGAAGTCGGTGACCTAACGAAAGAAGGAGCCGCCGAAGGCAGGTT GQ448468.1.1366 AGAGTTTGATCCTGGCTCAGGATGAACGCTGACAGAATGCTTAACACATGCAAGTATACTTGATCCTTCGGGTGAT GGTGGCGGACGGGTGAGTAACGCGTAAAGAACTTGCCCTGCAGTCTGGGACAACATTTGGAAACGAATGCTAATCC CGCATAAGCCCACAGCTCGGCATCGAGCAGAGGGAAAAGGAGTGATCTGCTTTGAGATGGCCTCGCGTCCGATTAG CTGGTTGGTGAGGTGACGGCCCATCAAGGCAACGATCGGTAGCCGGACTGAGAGGTTGAACGGCCACATTGGGATT GAGACACGGCCCTTACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGACCAAAAGTCTGATCCAGCAAT TCTGTGTGCACGATGAAGTTTTTCGGAATGTAAAGTGCTTTCAGTTGGGACGAAGTAAGTGACGGTACCAACAGAA GAAGCGACGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTA AAGCGCGTCTAGGCGGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCAA ACTAGAGTACTGGAGAGGTGGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATGG GGAAGCCAGCCCACTGGACAGATACTGACGCTAAAGCGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGT AGTCCACGCCGTAAACGATGATTACTAGGTGTTGGGGGTCGAACCTCAGCGCCCAAGCTAACGCGATAAGTAATCC GCCTGGGGAGTACGTACGCAAGTATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTT TAATTCGACGCAACGCGAGGAACCTTACCAGCGTTTGACATCCTAAGAAATTAGCAGAGATGCTTTTGTGCCCCTT CGGGGGAACTTAGTGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACG AGCGCAACCCCTTTCGTATGTTGCCATCATTAAGTTGGGCACTCATGCGATACTGCCTGCGATGAGCAGGAGGAAG GTGGGGATGACGTCAAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTACAATGGGTAGTACAGAGAGTC GCAAACCTGCGAGGGGGAGCTAATCTCAGAAAACTATTCTCAGTTCGGATTGTACTCTGCAACTCGAGTACATGAA GTTGGAATCGCTAGTAATCGCAAATCAGCTATGTTGCGGTGAATACGTTCTCGGGTCTTGTACACACCGCCCGT JN387556.1.1324 CGTAAGTAACCTGCCCTGTACACACGGATAACATACCGAAAGGTATGCTAATACGGGATAATATATTTTGATCGCA TGGTCGAGATATCAAAGCTCCGGCGGTACACCAGGGACCCCCGACAGAGGAGCTAGTTGGTAGTAATGTCACCAAG GCGACGATCAGAAGCCGAACTGAGAGGGGGATCCGCACATGACTGAGACACGGTCAAACTCCTACGGGAGGCAGCA GTGGGGAATATGCCAATGGGCGAAAGCTGATGCAGCACGCGCGTGAGCGATGAGGCTCGGGTCGTAAAGCTCGTCT CAAGGAAGATAATGACGGTACTTGAGGAGGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGG GCTAGCGTTATCCGGAATTACTGGGCGTAAAGGGTGCGTAGGCGGTCTTTCAAGTCAGGAGTGAAAGGCTACGGCT CAACCGTAGTAAGCTCTTGAAACTGTAAGACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAA TGCGTAGATATTAGGAGGAACACCAGTTGCGAAGGCGGCTCTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGT GGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTACTAGCTGTCGGAGGTTACCCCCT TCGGTGGCGCAGCTAACGCATTAAGTACTCCGCCTGGGAAGTACGCTCGCAAGAGTGAAACTCAAAGGAATTGACG GGGACCCGCACAAGTAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTAAGCTTGACATCCCA CTGACCCTTCCCTAATCGGAAGCTTCCCTTCGGGACAGTGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTC GTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGCCTTTAGTTGCCAGCATTAAGTTGGGCACTCTAGA GGGACTGCCAGGGATAACCCGGAGGAGTGGGGATGACGTCAAATCATCATGCCCTTATGCTAGGCTACACACGTGC TACAATGGGTGGTCAGAGGCCAGCCAGTCGTGAGGCCGAGCTATCCCATAAGCCATTCTCGTCCGGATTGTAGGCT GAACTCGCCTACATGAGCTGGAATTACAAGTATGCGATCGATGCTGCGTGATGCGTCCGGGTCTTGTACACACCGC CCGTCACACCATGGGAGTTGGGGGCGCCCGAAGCCGGATTGCTAACCTTTTGGAAGCGTCCGTCGAAGGTGAAACC AATAACTGGGGTGAAGTCGTAACAAGGTAACC OTUs in Table 4 JRPJ01000002.1034290.1035971 AGAGTTTGATCCTGGCTCAGAGTGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGATGAAACTTCTAGCT TGCTAGAAGTGGATTAGTGGCGCACGGGTGAGTAATGCATAGGTAACATGCCCTTTAGTCTGGGATAGCCACTGGA AACGGTGATTAATACTGGATACTCCCTACGGGGGAAAGGGGCTTTCAATAAAGAATTTCTCTTTTTAGTGTTTTGT GTTGTTGGCACAAAATTCTAGTATTTGGAATGAGAAATTGGTGTTGTGAAGCAATTTGTGCGGAGATTAGACTTAG TGTCTGTCGTGTCAGCAAATTGCGAACTCATCGATTTATCATCCAAAGACGAATTTTTTATTGAAAGCCTTCGCTA AAGGATTGGCCTATGTCCTATCAGCTTGTTGGTGAGGTAATGGCTCACCAAGGCTATGACGGGTATCCGGCCTGAG AGGGTGATCGGACACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTAGGGAATATTGCTCAAT GGGGGAAACCCTGAAGCAGCAACGCCGCGTGGAGGATGAAGGTTTTAGGATTGTAAACTCCTTTTGTAAGAGAAGA TTATGACGGTATCTTACGAATAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTT ACTCGGAATCACTGGGCGTAAAGAGCGCGTAGGCGGGTGGTCAAGTCAGATGTGAAATCCTGTAGCTTAACTACAG AACTGCATTTGAAACTGACCATCTAGAGTATGGGAGAGGTAGGTGGAATTCTTGGTGTAGGGGTAAAATCCGTAGA GATCAAGAGGAATACTCATTGCGAAGGCGACCTGCTGGAACATTACTGACGCTGATGCGCGAAAGCGTGGGGAGCA AACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGAATGCTAGTTGTTGTGAGGCTTGTCCTTGCAGTAA TGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATAGACGGGGACCC GCACAAGCGGTGGAGCATGTGGTTTAATTCGATGATACGCGAAGAACCTTACCTAGGCTTGACATTGATAGAATCT ACTAGAGATAGTGGAGTGCCCTTTTAGGGAGCTTGAAAACAGGTGCTGCACGGCTGTCGTCAGCTCGTGTCGTGAG ATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTCCTTAGTTGCTAGCAGTTTGGCTGAGCACTCTAAGGAGA CTGCCTTCGTAAGGAGGAGGAAGGTGAGGACGACGTCAAGTCATCATGGCCCTTACGCCTAGGGCTACACACGTGC TACAATGGGGTGCACAAAGAGATGCAATAGTGTGAGCTGGAGCCAATCTCTAAAACATCTCTCAGTTCGGATTGTA GTCTGCAACTCGACTACATGAAGCTGGAATCGCTAGTAATCGCAAATCAGCAATGTTGCGGTGAATACGTTCCCGG GTCTTGTACTCACCGCCCGTCACACCATGGGAGTTGTATTTGCCTTAAGTCGGAATGCTAAATTGGCTACCGCCCA CGGCAGATGCAGCGACTGGGGTGAAGTCGTAACAAGGTAACCGTAGGTGAACCTGCGGTTG New.ReferenceOTU45 TACGGAGGGTGCAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGTAGGCGGAAAGGCAAGCAAGATGTGAAA GACCTGGGCTCAACCTGGGTTGGTCATTTTGAACTACCTTTCTAGAGTATTGCAGAGGGAGATGGAATTTCAGGTG TAGCGGTGGAATGCGTAGATATCTGAAAGAACACCAGAGGCGAAGGCGGTCTCCTGGGCAAATACTGACGCTGAGG TGCGAAAGCGTGGGGAGCAAACAGG GQ006324.1.1342 GACGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGAAAGGCCCCAGCTCGCTGGGGTACTCGAGTGGCG AACGGGTGAGTAACACGTGGGTGATCTGCCTTGCACTCTGGGATAAGCTTGGGAAACTGGGTCTAATACCGGATAT GAACTGCCTTTAGTGTGGTGGTTGGAAAGTTTTTTCGGTGCAAGATGAGCTCGCGGCCTATCAGCTTGTTGGTGGG GTAATGGCCTACCAAGGCGTCGACGGGTAGCCGGCCTGAGAGGGTGTACGGCCACATTGGGACTGAGATACGGCCC AGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGGAAGCCTGATGCAGCGACGCCGCGTGGGGGA TGACGGCCTTCGGGTTGTAAACTCCTTTCGACAGGGACGAAGCTTTTTGTGACGGTACCTGTATAAGAAGCACCGG CTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTGTCCGGAATTACTGGGCGTAAAGAGCTCGT AGGTGGTTTGTCGCGTCGTCTGTGAAATTCCGGGGCTTAACTCCGGGCGTGCAGGCGATACGGGCATAACTTGAGT ACTGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCG GGTCTCTGGGCAGTAACTGACGCTGAGGAGCGAAAGCATGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCATG CCGTAAACGGTGGGCGCTAGGTGTGGGTTTCCTTCCACGGGATCCGTGCCGTAGCTAACGCATTAAGCGCCCCGCC TGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGTGGATTAA TTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATACACTGGATCGGGCTAGAGATAGTCTTTCCCTTTGTGG CTGGTGTACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAAC CCTTGTCTTATGTTGCCAGCATTTGGTTGGGGACTCATGAGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGAT GACGTCAAATCATCATGCCCCTTATGTCCAGGGCTTCACACATGCTACAATGGTCGGTACAACGCGCAGCGACACT GTGAGGTGGAGCGAATCGCTGAAAGCCGGCCTTAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGT CGCTAGTAATCGCAGATCAGCAATGCTGCGGTGAATACGTTCCCGGGCCT HK555938.1.1357 ACGGCACCCCTCTCCGGAGGGAAGCGAGTGGCGAACGGCTGAGTAACACGTGGAGAACCTGCCCCCTCCCCCGGGA TAGCCGCCCGAAAGGACGGGTAATACCGGATACCCCCGGGCGCCGCATGGCGCCCGGGCTAAAGCCCCGACGGGAG GGGATGGCTCCGCGGCCCATCAGGTAGACGGCGGGGTGACGGCCCACCGTGCCGACAACGGGTAGCCGGGTTGAGA GACCGACCGGCCAGATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTGCGCAATG GGGGGAACCCTGACGCAGCGACGCCGCGTGCGGGACGGAGGCCTTCGGGTCGTAAACCGCTTTCAGCAGGGAAGAG TCAAGACTGTACCTGCAGAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCGAGCGTT ATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGGCAGGCCGGGGGTCGAAGCGGGGGGCTCAACCCCCC GAAGCCCCCGGAACCTCCGCGGCTTGGGTCCGGTAGGGGAGGGTGGAACACCCGGTGTAGCGGTGGAATGCGCAGA TATCGGGTGGAACACCGGTGGCGAAGGCGGCCCTCTGGGCCGAGACCGACGCTGAGGCGCGAAAGCTGGGGGAGCG AACAGGATTAGATACCCTGGTAGTCCCAGCCGTAAACGATGGACGCTGGGTGTGGGGGGACGATCCCCCCGTGCCG CAGCCNACGCATTAAGCGTCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGC ACAAGCAGCGGAGCATGTGGCTTAATTCGAAGCAACGCGAAGAACCTTACGGCGCATCCCCCCGAGGCCCACGGGG GGTCCGCCGCGTGGGTCAGAGGAGCGCATACGGGAGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGT TAAGTCCCGCAACGAGCGCAACCCCCGCCGCGTGTTGCCATCGGGTGATGCCGGGAACCCACGCGGGACCGCCGCC GTCAAGGCGGAGGAGGGCGGGGACGACGTCAAGTCATCATGCCCCTTATGCCCTGGGCTGCACACGTGCTACAATG GCCGGTACAGAGGGATGCCACCCCGCGAGGGGGAGCGGATCCCGGAAAGCCGGCCCCAGTTCGGATTGGGGGCTGC AACCCGCCCCCATGAAGTCGGAGTTGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATGCGTTCCCGGGCCTTGT ACACACCGCCCGTCACACCACCCGAGTCGTCTGCACCCGAAGTCGCCGGCCCAACCGCAAGGGGG FJ957551.1.1489 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGGAGTTACTTTGAGAG CTTGCTTTCAAAGTAACTTAGCGGCGGACGGGTGAGTAACACGTAGGCAACCTGCCCCTTAGACTGGGATAACTAC CGGAAACGGTAGCTAATACCGGATAATTTCTTTTTTCTCCTGAAGGAAGAATGAAAGACGGAGCAATCTGTCACTG AGGGATGGGCCTGCGGCGCATTAGCTAGTTGGTGGGGTAACGGCCCACCAAGGCGACGATGCGTAGCCGACCTGAG AGGGTGATCGGCCACATTGGAACTGAGATACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAAT GGGGGAAACCCTGATGCAGCAACGCCGCGTGAGTGATGAAGGTCTTCGGATTGTAAAGCTCTGTCTTTAGGGACGA TAATGACGGTACCTAAGGAGGAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTT ATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATATTTAAGTGGGATGTGAAATACCCGAGCTTAACTTGGG AGCTGCATTCCAAACTGGATATCTAGAGTGCAGGAGAGGAGAATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGA GATTAGGAAGAACACCAGTGGCGAAGGCGATTCTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCA AACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATACCAGGTGTAGGGGCCCCAAGCCTCTGTGCCG CCGCTAACGCATTAAGTATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGC ACAAGCAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTAGACTTGACATGTCCTGAATTACC AGTAATGTGGGAAGTTCCTTCGGGAACAGGAACACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTT GGGTTAAGTCCCGCAACGAGCGCAACCCTTATTGTTAGTTGGTACCATTAAGTTGACCACTCTAGCGAGACTGCCC GGGTTAACCGGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGTCTAGGGCTACACACGTGCTACAA TGGCAAGTACAAAGAGAAGCAATACTGTGAAGTGGAGCAAAACTCAAAAACTTGTCTCAGTTCGGATTGTAGGCTG AAACTCGCCTACATGAAGCTGGAGTTGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGTCTTG TACACACCGCCCGTCACACCATGAGAGTTGGCAATACCCGAAGTCCGTAAGCTAACCGTAAGGAGGCAGCGGCCGA AGGTAGGGTCAGCGATTGGGGTGAAGTCGTAACAAGGTAACCAA FJ957494.1.1454 TGAGTTTGATCATGGCTCAGGACGAACGCTGGCGGCGTGCCTAACACATGCAAGTCGAGCGATGAAATTTTCTTCG GAAAATGGATTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTATAGAGAGGGATAGCCTTCCGAAAGG GAGATTAATACCTCATAATATCCTAGTATCGCATGATACATGGATTAAAGGAGCAATCCGCTATAGGATGGACCCG CGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCC ACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTG ATGCAGCAACGCCGCGTGAGTGATGACGGTCTTCGGATTGTAAAGCTCTGTCTTTAGGGACGATAATGACGGTACC TAAGGAGGAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTACT GGGCGTAAAGGGAGCGTAGGCGGATCTTTAAGTGGGATGTGAAATACTCGGGCTCAACCTGGGGGCTGCATTCCAA ACTGGGGATCTAGAGTACAGGAGGGGNGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAAC ACCAGTGGCGAAGGCGACTNTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGAT ACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTAGGGGGTGTCAACTCCCCCTGTGCCGCCGCTAACGC ATTAAGTATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGTAGCG GAGCATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCTAGACTTGACATCTTCTGCATTACCCTTAATCGGG GAAGTTCCTTCGGGGACAGAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTC CCGCAACGAGCGCAACCCTTAAGCTTAGTTGCCATCATTAAGTTGGGCACTCTAAGTTGACTGCCGGTGACAAACC GGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGTCTAGGGCTACACACGTGCTACAATGGCAAGTAC AAAGAGAAGCAATACTGTGAAGTGGAGCAAAACTCAAAAACTTGTCTCAGTTCGGATTGTAGGCTGAAACTCGCCT ACATGAAGCTGGAGTTGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGC CCGTCACACCATGAGAGTTGGCAATACCCGAAGTCCGTAAGCTAACCGTAAGGAGGCAGCGGCCGAAGGTAGGGTC AGCGATGGGG New.ReferenceOTU52 TACGTAGGTGGCGAGCGTTATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATGATTAAGTGGGATGTGAAA TACCCGGGCTCAACTTGGGTGCTGCATTCCAAACTGGTTATCTAGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTG TAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGACTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGG FM865905.1.1392 GGGAATCTCCAGGATCTGATTAGCGGCGGACGGGTGAGTACACGTGGGTAACCTGCCTCATAGAGTGGAATAGCCT TCCGAAAGGAAGATTAATACCGCATAACGTTGAAAGATGGCATCATCATTCAACCAAAGGAGCAATCCGCTATGAG ATGGACCCGCGGCGCATTAGCTAGTTGGTGGGGTAACGGCCTACCAAGGCGACGATGCGTAGCCGACCTGAGAGGG TGATCGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGG GAAACCCTGATGCAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTCTTTGGGGAAGATAAT GACGGTACCCAAGGAGGAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCGAGCGTTATCC GGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATGATTAAGTGGGATGTGAAATACCCGGGCTCAACTTGGGTGCT GCATTCCAAACTGGTTATCTAGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATT AGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACA GGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTGGGGGTTTCAACACCTCCGTGCCGCCG CTAACGCATTAAGTATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTNAAAGGAATTGACGGGGATCNNCACN AGTAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTACACTTGACATCCCTTGCATTACTCTT AATCGAGGAAATCTCTTCGGGGACAAGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGG TTAAGTCCCGNAACGAGGGGAACCNTTGTCGTTAGTTACTACCATTAAGTTGAGGACTNTAGNGAGACNGCTGGGT TAACNAGGAGGAAGGTGGGGATGACTCAATCTCTGGNCNTTATGTGTAGGGNTACACACGTGCTACAATGGCTGGT ACAGAGAGATGCATACCGGGAGGTGGANTCAATTTAAAAACAGTNTCNTTCGGATTGTAGGNTGAANTNNCCTACT GAAGNTGGAGTTANTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGTCTTGTACACNCCNCCCGT CACNCCATGAGAGTTGGCAATACCCGAAGTCCGTGAGCTAACCGCAAGGAGGCAGCGGCCGAAGGTAGGGTCAGCG ATTGGGGTGAAGTCGTAACAGGNA GQ016239.1.1362 GATGAACGCTGGCGGCATGCCTAATACATGCAAGTCGAACGGAGCGAATATGGAAGCTTGCTTCCGTAAGAGCTCA GTGGCGAACGGGTGAGTAACACGTAGGTAACCTGCCCATGTGCCCGGGATAACTGCTGGAAACGGTAGCTAAAACC GGATAGGTGAATAGGAGGCATCTCTTATTCATTAAAGGACCTGTAAGGGTGCGAACATGGATGGACCTGCGGCGCA TTAGCTGGTTGGAGTGGTAACGGCACACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGCGAACGGCCACATTGG GACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATTTTCGTCAATGGGGGGAACCCTGAACGAGC AATGCCGCGTGAGTGAAGAAGGTCTTCGGATCGTAAAGCTCTGTTGTAAGTGAAGAACGGTCAGTAGAGGAAATGA TACTGAAGTGACGGTAGCTTACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCGA GCGTTATCCGGAATCATTGGGCGTAAAGGGTGCGCAGGTGGTACATTAAGTCCGAAGTAAAAGGCAGCAGCTCAAC TGCTGTTGGCTTTGGAAACTGGTGAACTGGAGTGCAGGAGAGGGCGATGGAATTCCATGTGTAGCGGTAAAATGCG TAGATATATGGAGGAACACCAGTGGCGAAGGCGGTCGCCTGGCCTGCAACTGACACTGAGGCACGAAAGCGTGGGG AGCAAATAGGATTAGATACCCTAGTAGTCCACGCCGTAAACGATGAGAACTAAGTGTTGGGGAGACTCAGTGCTGC AGTTAACGCAATAAGTTCTCCGCCTGGGGAGTATGCACGCAAGTGTGAAACTCAAAGGAATTGACGGGGGCCCGCA CAAGCGGTGGAGTATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATGGATGTAAATGTTC TAGAGATAGAAAGATAGCTATACATCACACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTT AAGTCCCGCAACGAGCGCAACCCTTATCGCATGTTACCAGTATTGAGTTAGGGACTCATGCGAGACTGCCGGTGAC AAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGGCCTGGGCTACACACGTACTACAATGGCG GCTACAAAGAGAAGCGAACCTGCGAGGGGGAGCGGAACTCATAAAGGCCGTCTCAGTTCGGATTGGAGTCTGCAAC TCGACTCCATGAAGTCGGAATCGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCTCGGGCCT HG798451.1.1400 CTTGTGTCACCAACCATAGGGAGGGGGAAAACATGGAAACGGGGTTCATACCGCATAACTTTTTTAGCCCAATGCA TAAGAAGAAAGGCCTTTCGGGTTTCGGTAAAGGAGGCCCCCGCGGCTCTTATAGTGTGTGTGGAAGTAACCGCTTC CACAAGGCCCAGGTTTCATACCCGACTGGAGAGTGTGTTCGCCACACTGGGGAAAGGACCCCCGGCCCAGTCTCTC TAGGGGAGGCAGCAGTAGGAATTTTCGGCAAAGGAAAAAATTTCTGACCGAACAACGCCGGTTGAATGAAGAAGTT TTTCGGATCGAAAAACTCTGTTGTTAGAGAAGAACAAGGACGTTAGTAACTGAACGTCCCCTGACGGTATCTAACC AGAAAGCCACGGCTAATTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCG TAAAGCGAGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGG GAGACTTGAGTGCAGAAGAGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCAG TGGCGAAGGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCT GGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCAAACGCATTAA GCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCA TGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTTTGACCACTCTAGAGATAGAGCTTT CCCTTCGGGGACAAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA ACGAGCGCAACCCTTATTGTTAGTTGCCATCATTTAGTTGGGCACTCTAGCGAGACTGCCGGTGACAAACCGGAGG AAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGAAGTACAACGA GTCGCTAGACCGCGAGGTCATGCAAATCTCTTAAAGCTTCTCTCAGTTCGGATTGCAGGCTGCAACTCGCCTGCAT GAAGCCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGT CACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTTGGAGCCAGCCGCCTAAGGTGGGATAGAT GATTGGGGTGAAGTCGTAACCAACGTATGCC EU461791.1.1414 AGAGTTTGATCCTGGCTCAGGACTAACGCTGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGACTTTAGCT TGCTAAAGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGATAACTATTGGA AACGATAGCTAATACCGCATAACAGCATTTAACCCATGTTAGATGCTTGAAAGGAGCAATTGCTTCACTAGTAGAT GGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGACCTGAGAGGGTG ATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGC AACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGT GTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTA ATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAA AGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTG TAGCGGTGAAATGCGTAGATATATGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGAAAGGTGTTAGG CCCTTTCCGGGGCTTAGTTGCTGCACGCTAACTGCATTATGACACTCCGCCAGGGGAGTACGACCGCTAGGTTGAA ACTCAAAGGAGTTGACGGGGGCCAGCACAACCGGTGGAGCATGTGGTTGAATTGGAAGCAACGCGAAGAGCCTTAC CAGGTCTTGACATCCCGACGCTATTCCTAGAGATAGGAAGTTTCTTCGGGACATTCGGTGGCAGGTGGTGCATGGT AGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATACAT TAAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCT TATGACCTGGGCTACACACGACGCTACAATGGTTGGTACAACGAGTCGCGAGTCGGTGACGGCAAGCAAATCTCTT AAAGCCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGC ACGCCGCGGTGAATACGTTCCCGGGCCTTGCACTCACCGCCCGTCA GU303759.1.1517 AGAGTTTGATCATGGCTCAGGACGAACGCCGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGACTTTAGCT TGCTAAAGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGATAACTATTGGA AACGATAGCTAATACCGTATAACAGCATTTAACACATGTTAGATGCTTGAAAGGAGCAATTGCTTCACTAGTAGAT GGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGACCTGAGAGGGTG ATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGC AACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGT GTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTA ATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAA AGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTG TAGCGGTGAAATGCGTAGATATATGGARGGAAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGA GGCTCGAGAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAGCGATGAGTGCTAGGTGTT AGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAA CTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACC AGGTCTTGACATCCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACATCGGTGACAGGTGGTGCATGGTTG TCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCATTAA GTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTAT GACCTGGGCTACACACGTGCTACAATGGCGGTCAACAGAGGGAAGCAATACTGTGAAGTGGAGCAAACCCCTAAAA GCCGTCCCAGTTCGGATTGCAGGCTGCAACCCGCCTGTATGAAGTTGGAATCGCTAGTAATCGCGGATCAGCATGC CGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTCGGGAACACCCGAAGTCCGTAG CCTAACTTTCACGAGGGGGCGCGGCCGAAGGTGGGTTCGATAATTGGGGTGAAGTCGTAACAAGGTAACCGTA New.ReferenceOTU114 TACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAAA GGCAGTGGCTTAACCATTTTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTGT AGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGGC TCGAAAGCGTGGGGAGCAAACAGG AB506154.1.1541 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGAAAGGAGCT TGCTTCTTTTGGATGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTTGTAGCGGGGGATAACTATTGGA AACGATAGCTAATACCGCATAACAGCTTTTGACACATGTTAGAAGCTTGAAAGATGCAATTGCATCACTACGAGAT GGACCTGCGTTGTATTAGCTAGTAGGTAGGGTAACGGCCTACCTAGGCGACGATACATAGCCGACCTGAGAGGGTG ATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGC AACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGT GTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTG ATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAA AGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAAACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTG TAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTAGG CCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGTATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTC AAAGGAATTGACGGGGGCCCGCACAAGCAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTAG ACTTGACATCTCCTGCATTACTCTTAATCGAGGAAGTCCCTTCGGGGACAGGATGACAGGTGGTGCATGGTTGTCG TCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTGTTAGTTGCCATCATTAAGTT GGGCACTCTAGCGAGACTGCCCGGGTTAACCGGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGTC TAGGGCTACACACGTGCTACAATGGTCGGTACAATAAGACGCAAGCCCGCGAGGGGGAGCAAAACTGGAAAACCGA TCTCAGTTCGGATTGTAGGCTGAAACTCGCCTACATGAAGCTGGAGTTGCTAGTAATCGCGAATCAGCATGTCGCG GTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTGGCAATACCCAAAGTACGTGATCTA ACCCGCAAGGGAGGAAGCGTCCTAAGGTAGGGTCAGCGATTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAAC CTGCGGCTG EU774370.1.1398 AGAGTTTGCTCTTGGGTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAGCGAGAAAAGTTCTTCGG AGCTTTTCTAGCGGCGGACGGGTGAGTAACACGTGGGCAACCTGCCTCATAGAGGGGAATAGCCTTCCGAAAGGAA GATTAATACCGCATAACATTGTTGAAAGGCATCTTTTAACAATCAAAGGAGCAATCCGCTATGAGATGGGCCCGCG GCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCAC ATTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGAT GCAGCGACGCCGCGTGAGTGAAGAAGTATTTCGGTATGTAAAGCTCTGTTGTAAGAGAAGAACGTGTGTGAGAGTG GAAAGTTCACACAGTGACGGTAACTTACCAGAGAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGG TCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAAAGGCAGTGG CTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGA AATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGC GTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTAGGCCCTTTCCG GGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATT GACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACAT CCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACATCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGT GTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCATTAAGTTGGGCACTCT AGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTAC ACACGTGCTACAATGGTTGGTACAACGAGTCGCGAGTCGGTGACGGCAAGCAAATCTCTTAAAGCCAATCTCAGTT CGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATAC GTTCCCGGGCCTTGCACTCACCGCCCGTCA HK557089.3.1395 AGACTTTAGCTTGCTAAAGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGA TAACTATTGGAAACGATAGCTAATACCGCATAACAGCATTTAACCCATGTTAGATGCTTGAAAGGAGCAATTGCTT CACTAGTAGATGGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGAC CTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCG GCAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGA GAAGAACGTGTGTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAG TCTGAAGTTAAAGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGG AATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAAC TGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGC TAGGTGTTAGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAA GGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGA ACCTTACCAGGTCTTGACATCCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACTGTGAGACTTGAGGGCA GAAGGGTAGAGTGCACTTGTATGGGGAGCTGTGGAATGCGTTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCC ATCATTAAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATG CCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTACAACGAGTCGCGAGTCGGTGACGGCAAGCAAATC TCTTAAAGCCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGAT CAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAA GTCGGTGAGGTANCCTTTTAGGAGC HQ807346.1.1456 TTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGACTTTAGCTTGCTAAAGTTGGAAG AGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGATAACTATTGGAAACGATAGCTAATAC CGCATAACAGCATTTAACCCATGTTAGATGCTTGAAAGGAGCAATTGCTTCACTAGTAGATGGACCTGCGTTGTAT TAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGACCTGAGAGGGTGATCGGCCACACTGGG ACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGCAACCCTGACCGAGCA ACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGTGTGAGAGTGGAAAGT TCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGA GCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAAAGGCAGTGGCTTAAC CATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCG TAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGG AGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTAGGCCCTTTCCGGGGCTT AGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGG GGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTTTG ACCACTCTAGAGATAGAGCTTCCCCTTCGGGGGCAAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTG AGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTGTTAGTTGCCATCATTTAGTTGGGCACTCTAGCGAG ACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGT GCTACAATGGGAAGTACAACGAGTTGCGAAGTCGCGAGGCTAAGCTAATCTCTTAAAGCTTCTCTCAGTTCGGATT GTAGGCTGCAACTCGCCTACATGAAGCCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCC GGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTAGGAGCC AGCCGCCTAAGG HQ748204.1.1442 CTAATACATGCGAGGAGAACGCTGAAGACTTTCTTTTGCTATAGTTGGGAGAGTTGCTAACGGGTGAGTAACGCGT AGGTGACCTGCCTACTAGCGGGGGATAACTATTGCAAACGATAGCTAATACCGCATAACAGCCTTTAACCCATGTT AGATGCTTGAAAGGAGCAATTGCTTCACTAGTAGATGGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTC ACCAAGGCGACGATACATAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCATACTCCTAC GGGAGGCACCAGTAGGGAATCTTCGGGAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTT CGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGTGTGAGAGTGGAAAGTTCACACTGTGACGGTAACTTACCAG AAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTA AAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAAAGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAG ACTTGAGTGCATAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGG CGAAAGCGGCTCTCTGGTCTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGT AGTCCACGCTGTAAACGATGAGTGGTAGGTGTTAGGCCCTTTCTGGGGTTTAGTGCCGCAGATTACGCATTAAGCC ATTCGCCTGGGGAGTACGACCGCAAGGTTGAAACTTAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGT GGTTTAATTAGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCGATGCTATTCTTAGAGATAGGAAGTTTC TTCGGAACATCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGAGAGATGTTGGGTTAAGTCCCTCAACG AGCGCAACCCCTATTTTTATTTGCCATCATTAAGTTGGGCAATCTAGCGAGACTGCCGGTAATAAACCGGAGGAAG GTGGGGATGACGTCAAATCATCATGCTCCTTATGTCATGGGGTACACACGTGGTACAATGGTTGGTACAACGAGTC GCGAGTTGGTGAAGGCAAGCAAATCTCTTAAAGCCAATATCAGTTCGGATTGTAGGCTGCAAATAGCCTACATGTA GTCGGAATTGTTAGTAATCGGGGATCAGCACTCCGCGGTGAATACGTTTCCGGGCCTTGTACACCCCGCCCGTCTA CACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACTCTTTTAGGAGCCAGCCGCCTAAGGTGGGATAGA GU179917.1.1382 AGAGTTTGATTATGGCTCAGGATGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGCGAGCAGCAATGCTC GAGTGGCGAACGGGTGAGTAATACATAAGTAACCTGCCCTAGACAGGGGGATAACTGCTGGAAACGGCAGCTAAGA CCGCATAGGTATGGACACTGCATGGTGACCATATTAAAAGTGCCAAGGCACTGGTAGAGGATGGACTTATGGCGCA TTAGCTGGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGACCGGCCACACTGG GACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATTTTCGGCAATGGGGGGAACCCTGACCGAGC AACGCCGCGTGAAGGAAGAAGGAATTCGTTCTGTAAACTTCTGTTATAAAGGAAGAACGGCGGATATAGGGAATGA TATCCGAGTGACGGTACTTTATGAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCGA GCGTTATCCGGAATTATTGGGCGTAAAGAGGGAGCAGGCGGCGGCAGAGGTCTGTGGTGAAAGACTGAAGCTTAAC TTCAGTAAGCCATAGAAACCGGGCTGCTAGAGTGCAGGAGAGGATCGTGGAATTCCATGTGTAGCGGTGAAATGCG TAGATATATGGAGGAACACCAGTGGCGAAGGCGACGGTCTGGCCTGTAACTGACGCTCATTCCCGAAAGCGTGGGG AGCAAATAGGATTAGATACCCTAGTAGTCCACGCCGTAAACGATGAGTACTAAGTGTTGGGAGTCAAATTTCAGTG CTGCAGTTAACGCAATAAGTACTCCGCCTGAGTAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGGCC CGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCAGGGCTTAAATGTGACTGACAG GTCCGGAAACGGACTTTTCTTCGGACAGTTACAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCAGG TTAAGTCCTATAACGAGCGCAACCCCTGTCGCTAGTTGCCAGCGAGTAATGTCGGGAACTCTAGCGAGACTGCCAG TGCAAACTGCGAGGAAGGTGGGGATGACGTCAAATCATCACGGCCCTTACGCCCTGGGCTACACACGTGCTACAAT GGCCGGTACAGAGAGCAGCCACCCCGCGAGGGGGAGCGAATCTACAAAACCGGTCACAGTTCGGATCGGAGTCTGC AACTCGACTCCGTGAAGCTGGAATCGCTAGTAATCGGATATCAGCCATGATCCGGTGAATACGTTCCCGGGCCTTG TACACACCCCCGTC GQ448336.1.1418 AGAGTTTGATCATGGCTCAGGACGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAGAAGAGATGAGAAG CTTGCTTCTTATCTCTTCGAGTGGCAAACGGGTGAGTAACGCGTAAGCAACCTGCCCTTCAGATGGGGACAACAGC TGGAAACGGCTGCTAATACCGAATACGTTCTTTTTGTCGCATGGCAGAGGGAAGAAAGGGAGGCTCTTCGGAGCTT TCGCTGAAGGAGGGGCTTGCGTCTGATTAGCTAGTTGGAGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCCGG TCTGAGAGGATGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTC CGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAACGATGACGGCCTTCGGGTTGTAAAGTTCTGTTATACG GGACGAATGGCGTAGCGGTCAATACCCGTTACGAGTGACGGTACCGTAAGAGAAAGCCACGGCTAACTACGTGCCA GCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCGCGCAGGCGGCGTCGTAA GTCGGTCTTAAAAGTGCGGGGCTTAACCCCGTGAGGGGACCGAAACTGCGATGCTAGAGTATCGGAGAGGAAAGCG GAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAAGCGGCTTTCTGGACGACAA CTGACGCTGAGGCGCGAAAGCCAGGGGAGCAAACGGGATTAGATACCCCGGTAGTCCTGGCCGTAAACGATGGATA CTAGGTGTAGGAGGTATCGACCCCTTCTGTGCCGGAGTTAACGCAATAAGTATCCCGCCTGGGGAGTACGGCCGCA AGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAACATGTGGTTTAATTCGATGATACGCGAGG AACCTTACCCGGGCTTAAATTGCAGTGGAATGATGTGGAAACATGTCAGTGAGCAATCACCGCTGTGAAGGTGCTG CATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGCCATAACGAGCGCAACCCTTATCTTCAGTTACT AACAGGTCATGCTGAGGACTCTGGAGAGACTGCCGTCGTAAGATGTGAGGAAGGTGGGGATGACGTCAAATCAGCA CGGCCCTTACGTCCGGGGCTACACACGTGTTACAATGGGGGGTACAGAGGGCCGCTACCACGCGAGTGGATGCCAA TCCCAAAAACCTCTCTCAGTTCGGACTGGAGTCTGCAACCCGACTCCACGAAGCTGGATTCGCTAGTAATCGCGCA TCAGCCACGGCGCGGTGAATACGTTCCCGGGCCTTGCACTCACCGCCCGT DQ804865.1.1390 AGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGGAAACTTTTCATTG AAGCTTCGGCAGATTTGGTCTGTTTCTAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTTATACAGGGGG ATAACAACCAGAAATGGTTGCTAATACCGCATAAGCGCACAGGACCGCATGGTCCGGTGTGAAAAACTCCGGTGGT ATAAGATGGACCCGCGTTGGATTAGCTAGTTGGCAGGGTAACGGCCTACCAAGGCGACGATCCATAGCCGGCCTGA GAGGGTGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAA TGGGGGAAACCCTGATGCAGCGACGCCGCGTGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAG ATAGTGACGGTACCTGACTAAGAAGCCCCGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGGTTCAAGCGT TATCCGGATTTACTGGGTGTAAAGGGTGAGTAGGCGGTTATGCAAGTCATATGTGAAATGTCGGGGCTCAACTCCG GCCTGCATAAGAAACTGTATAACTAGAGTGCAGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGA TATTAGGAAGAACACCGGTGGCGAAGGCGGCTTGCTGGACTGTTACTGACGCTGAGTCACGAAAGCGTGGGGAGCA AACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTCGGGTGGCAAAGCCATTCGGTGC CGCAGCAAACGCAATAAGTATTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCC GCACAAGCGGTGGAGTATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATGGATATAAATG TTCTAGAGATAGAAAGATAGCTATATATCACACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGG GTTAAGTCCCGCAACGAGCGCAACCCTTGTCTTCTGTTACCAGCATTGAGTTGGGGACTCAGGAGAGACTGCCGGT GACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGGCCTGGGCTACACACGTACTACAATG GCGCCTACAAAGAGCAGCGACACCGCGAGGTGAAGCGAATCTCATAAAGGGCGTCTCAGTTCGGATTGAAGTCTGC AACTCGACTTCATGAAGTCGGAATCGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCCCGGGTCTTGT ACTCACCGCCCGTCA GQ491757.1.1361 GATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAGCGAAGCACTTAAGTGGATCTCTTCGGATTGAAACTTA TTTGACTGAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGGGGGATAACAGTTAGAAATGGCT GCTAATACCGCATAAGCGCACAGGACCGCATGGTCTGGTGTGAAAAACTCCGGTGGTATGAGATGGACCCGCGTCT GATTAGCTAGTTGGAGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCCGGCCTGAGAGGGTGAACGGCCACATT GGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCA GCGACGCCGCGTGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGACGGTACCTGACT AAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACTGGGTG TAAAGGGAGCGTAGACGGAAGAGCAAGTCTGATGTGAAAGGCTGGGGCTTAACCCCAGGACTGCATTGGAAACTGT TTTTCTAGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCGG TGGCGAAGGCGGCTTACTGGACGACCACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCT GGTAGTCCACGCCGTAAACCGATGAATAATAGGTGTCGGGGAACAATAGTTCTTTGGTGCCGCAGCAAAACGCATT AAGTATTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAG CATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGCTCTTGACATCCCACTGACCGGACAGTAATGTGTCC TTTTCTTCTGAACAGTGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCG CAACGAGCGCAACCCTCGTCTTTAGTAGCCAGCAGTCCGGCTGGGCACTCTAGAGAGACTGCCAGGGATAACCTGG AGGAAGGCGGGGAGGACGTCAAATCATCATGCCCCTTACGAGCAGGGCTACACACGTGCTACAATGGCGTAAACAA AGGGAAGCGACCCCGTGAAGGTGAGCAAATCTCAAAAATAACGTCTCAGTTCGGATTGTAGTCTGCAACTCGACTA CATGAAGCTGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATAAAAGGCCGGGTCTTGCACA New.ReferenceOTU56 TACGGAAGGTCCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCGGACTCTTAAGTCAGTTGTGAAA TACGGCGGCTCAACCGTCGGACTGCAGTTGATACTGGGAGTCTTGAGTACACGCAGAGATACTGGAATTCATGGTG TAGCGGTGAAATGCTCAGATATCATGAGGAACTCCGATCGCGAAGGCAGGTATCTGGAGTGTAACTGACGCTGAGG CTCGAAAGTGCGGGTATCAAACAGG KF842598.1.1394 AGAGTTTGATCCTGGCTCAGGATGAACGCTAGCGACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGATTTGTA GCAATACAGATTGATGGCGACCGGCGCACGGGTGAGTAACGCGTATGCAACTTACCTATCAGAGGGGGATAGCCCG GCGAAAGTCGGATTAATACCCCATAAAACAGGGGTCCCGCATGGGAATATTTGTTAAAGATTCATCGCTGATAGAT AGGCATGCGTTCCATTAGGCAGTTGGCGGGGTAACGGCCCACCAAACCGACGATGGATAGGGGTTCTGAGAGGAAG GTCCCCCACATTGGTACTGAGACACGGACCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGCCGA GAGGCTGAACCAGCCAAGTCGCGTGAAGGAAGAAGGATCTATGGTCTGTAAACTTCTTTTATAGGGGAATAAAGTG GAGGACGTGTCCTTTTTTGTATGTACCCTATGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGG AGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGTGGTGATTTAAGTCAGCGGTGAAAGTTTG TGGCTCAACCATAAAATTGCCGTTGAAACTGGGTTACTTGAGTGTGTTTGAGGTAGGCGGAATGCGTGGTGTAGCG GTGAAATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCTTACTAAACCATAACTGACACTGAAGCACGA AAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCAGTAAACGATGATTACTAGGAGTTTGCGATAC AATGTAAGCTCTACAGCGAAAGCGTTAAGTAATCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATT GACGGGGGCCCGCACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGTTTGAACG TAGTCTGACCGGAATGGAAACACTCCTTCTAGCAATAGCAGATTACAAGGTGCTGCATGGTTGCCTCAACTCCGGC CCGGAAGGTCCGGCTTAATTGCCATAACAAGCGCACCCTTTTACCAAGGTTCAAACAGGTGAAGCTTGAAGACTCT GTGGAACCTCCCCCCTAACCTGTGAGAAGAAGTGGGGATACACTCAATAAACCACGGCCCTTAATCCCGGGGGGAA CACTGGTTACAATGGGTTGGGAAAGGGGGCTTCCTGGCGACAGGATGCTAATCTCCAAACCATGTCTCAGTTCGGA TCGGAGTCTGCAACTCGACTCCGTGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTT CCCGGGCCTTGTACACACCGCCCGTC HQ802052.1.1445 TACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGATTGAAGCTTGCTTCAATTGATGGCGACCGGCGCACGGGT GAGTAACACGTATCCAACCTTCCGTACACTCAGGGATAGCCTTTCGAAAGAAAGATTAATACCTGATGGTATCTTA AGCACACATGTAATTAAGATTAAAGATTTATCGGTGTACGATGGGGATGCGTTCCATTAGGTAGTAGGCGGGGTAA CGGCCCACCTAGCCTACGATGGATGGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGACACGGTCCAAAC TCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGACGAGAGTCTGAACCAGCCAAGTAGCGTGAAGGATGAA GGTCCTACGGATTGTAAACTTCTTTTATAAGGGAATAAAACCTCCCACGTGTGGGAGCTTGTATGTACCTTATGAA TAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATGCGAGCGTTATCCGGATTTATTGGGTTTA AAGGGAGCGCAGACGGGTCGTTAAGTCAGCTGTGAAAGTTTGGGGCTCAACCTTAAAATTGCAGTTGATACTGGCG TCCTTGAGTGCGGTTGAGGTGTGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATT GCGAAGGCAGCACACTAAGCCGTAACTGACGTTCATGCTCGAAAGTGTGGGTATCAAACAGGATTAGATACCCTGG TAGTCCACACAGTAAACGATGAATACTCGCTGTTTGCGATATACAGTAAGCGGCCAAGCGAAAGCATTAAGTATTC CACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGAGGAACATGTGGT TTAATTCGATGATACGCGAGGAACCTTACCCGGGCTTAAATTGCATTTGAATATATTGGAAACAGTATAGCCGTAA GGCAAATGTGAAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGCCATAACGAGCGCA ACCCTTATCTTCAGTTACTAACAGGTCATGCTGAGGACTCTGGAGAGACTGCCGTCGTAAGATGTGAGGAAGGTGG GGATGACGTCAAATCAGCACGGCCCTTACGTCCGGGGCTACACACGTGTTACAATGGGGGGTACAGAAGGCCGCTA CCTGGTGACAGGATGCTAATCCCAAAAGCCTCTCTCAGTTCGGATCGAAGTCTGCAACCCGACTTCGTGAAGCTGG ATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCAAGCCA TGAAAGCCGGGGGTACCTGAAGTACGTAACCGCAAGGAGCGTCCTAGGGTAAAACTGGTAATTGGGGCTAAGTCAT A GX182404.8.1529 AGAGTTTGATCATGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAACGGTAACAGGAAGAAGC TTGCTTCTTTGCTGACGAGTGGCGGACGGGTGAGTAATGTCTGGGAAACTGCCTGATGGAGGGGGATAACTACTGG AAACGGTAGCTAATACCGCATAACGTCGCAAGACCAAAGAGGGGGACCTTCGGGCCTCTTGCCATCGGATGTGCCC AGATGGGATTAGCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACGATCCCTAGCTGGTCTGAGAGGATGACCAGC CACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCT GATGCAGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGAGTAAAGT TAATACCTTTGCTCATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGA GGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTTGTTAAGTCAGATGTGAAATCCCCG GGCTCAACCTGGGAACTGCATCTGATACTGGCAAGCTTGAGTCTCGTAGAGGGGGGTAGAATTCCAGGTGTAGCGG TGAAATGCGTAGAGATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACGAAGACTGACGCTCAGGTGCGAA AGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTT GAGGCGTGGCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAA TTGACGGGGGCCCGCACAAGCGGCGGAGCATGTGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGGTTTGAC ATGCACAGGACGCGTCTAGAGATAGGCGTTCCCTTGTGGCCTGTGTGCAGGTGGTGCATGGCTGTCGTCAGCTCGT GTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCTCATGTTGCCAGCACGTAATGGTGGGGACT CGTGAGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGTCCAGGGCT TCACACATGCTACAATGGCCGGTACAAAGGGCTGCGATGCCGCGAGGTTAAGCGAATCCTTAAAAGCCGGTCTCAG TTCGGATCGGGGTCTGCAACTCGACCCCGTGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCGGTGAA TACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCATGAAAGTCGGTAACACCCGAAGCCAGTGGCCTAACCCT CGGGAGGGAGCTGTCGAAGGTGGGATCGGCGATTGGGACGAAGTCGTAACAAGGTAACCGTAGGGGAACCTGCGGT TG FJ950694.1.1472 CGCCCTGATTGACGGCTATACACATGCAAGTCGAACGGTAACAGGAAACAGCTTGCTTCTTTGCTGACGAGTGGCG GACGGGTGAGTAATGTCTGGGAAACTGCCTGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAAC GTCGCAAGACCAAAGAGGGGGACCTTCGGGCCTCTTGCCATCGGATGTGCCCAGATGGGATTAGCTAGTAGGTGGG GTAACGGCTCCATCCCTAGGCGAGCCGAATCCTTAGCCTGGTCTGAGAGGAATGACCAGCCACACTGGGACTGAGA ACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCG CGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGAGTAAAGTTAATACCCTTTGCTC ATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTA ATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTTGTTAAGTCAGATGTGAAATCCCCGGGCTCAACCTGGGA ACTGCATCTGATACTGGCAAGCTTGAGTCTCGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAG ATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACGAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAA ACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTGGCTTCC GGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCG CACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGTCTTGACATCCACGGGAAGTT TTCAGAGATGAGAATGTGCCTTCGGGAACCGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATG TTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCAGCGGTCCGGCCGGGAACTCAAAGGAGACTG CCAGTGATAAACTGGAGGAAGGTGGGGATGACGTCCAGGTCATCATGGCCCTTACGAACCAGGGCTACACACGTGC CTACAATGGACGCATCCAAAGAGAGAGCGAACCCTGCCCGCGAGAGCAAGCGGACCTCATAAAGTGCGTCGTAGTC CGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTGGATCAGAATGCCACGGTGAATAC GTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCAAAAGAAGTAGGTAGCTTAACCTTCGG GAGGGCGCTTACCACTTTGGATGCGAGG GQ448506.1.1374 AGAGTTTGATCATGGCTCAGGATGAACGCTAGCTACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGGTCTTAG CTTGCTAAGGCCGATGGCGACCGGCGCACGGGTGAGTAACACGTATCCAACCTGCCGTCTACTCTTGGACAGCCTT CTGAAAGGAAGATTAATACAAGATGGCATCATGAGTCCGCATGTTCACATGATTAAAGGTATTCCGGTAGACGATG GGGATGCGTTCCATTAGATAGTAGGCGGGGTAACGGCCCACCTAGTCTTCGATGGGTAGGGGTTCTGAGAGGAAGG TCCCCCACATTGGAACTGAGACACGGCCCAAACTCATACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAA ACCCTGATGCAGCGACGCCGCGTGAAGGATGAAGTATTTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGAC GGTACCTGACTAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGA TTTACTGGGTGTAAAGGGAGCGTAGACGGCAGTGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCT TTGGAAACTGTGCAGCTAGAGTGTCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGG AGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGA TTAGATACCCTGGTAGTCCACGCCGTAAACGATGACTACTAGGTGTCGGGGAGCAAAGCTCTTCGGTGCCGCAGCC AACGCAATAAGTAGTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAG CGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGCTCTTGACATCCCTCTGACCGCTCTTTA ATCGGAGCTTTCCTTCGGGACAGAGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTT AAGTCCCGCAACGAGCGCAACCCTTATGGTCAGTTACTACGCAAGAGGACTCTGGCCAGACTGCCGTTGACAAAAC GGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCTTTATGACTTGGGCTACACACGTACTACAATGGCGTTAAA CAAAGAGAAGCGAGACCGCGAGGTGGAGCAAAACTCGGAAACAACGTCCCAGTTCGGACTGCAGGCTGCAACTCGC CTGCACGAAGTCGGAATTGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCCCGGGCCTTGCACTCACC GCCCGT HQ802983.1.1440 TAAGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCCTATGAAGCGCTTAAACGGATTTCTTCGGATTGAAGT TTTTGTGACTGAGTGGCGGACGGGTGAGTAACGCGTGGGTAACTTGCCTCATACAGGGGGATAACAGTTAGAAATG ACTGCTAATACCGCATAAGCGCACAGTGCTGCATGGCACAGTGTGAAAAACTCCGGTGGTATGAGATGGACCCGCG TCTGATTAGCTAGTTGGTGGGGTAACGGCCTACCAAGGCGACGATCAGTAGCCGGCCTGAGAGGGTGAACGGCCAC ATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGAT GCAGCGACGCCGCGTGAGCGAAGAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGGAAGAAAATGACGGTACCTG ACTAAGAAGCACCGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGGTGCAAGCGTTATCCGGATTTACTGG GTGTAAAGGGAGCGTAGACGGTTGTGTAAGTCTGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTGGAAAC TATGTAACTAGAGTGTCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACAC CAGTGGCGAAGGCGGCTTACTGGACGATCACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATAC CCTGGTAGTCCACGCCGTAAACGATGACTACTAGGTGTCGGGGCCCATAAGGGCTTCGGTGCCGCAGCAAACGCAA TAAGTATTCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGA GCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGGTCTTGACATCCCACTGACCGGACAGTAATGTGTC CTTTCCTCCGGGACAGTGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCC GCAACGAGCGCAACCCCTATCCTTAGTAGCCAGCAGTAAGATGGGCACTCTAGGGAGACTGCCAGGGATAACCTGG AGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACTTGGGCTACACACGTGCTACAATGGCGTAAACAA AGTGAAGCGAAGTCGTGAGGCCAAGCAAATCACAAAAATAACGTCTCAGTTCGGATTGTAGTCTGCAACTCGACTA CAAGAAGCTGGAATCGCTAGTAATCGCAGATCAGAATGCTGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCC CGTCACACCATGGGAGTCGAAAATGCCCGAAGTCGGTGACCTAACGAAAGAAGGAGCCGCCGAAGGCAGGTT DQ793824.1.1370 ATGAACGCTGGCGGCGTGCCTAACACATGCAAGTCGAACGAAGCGCTTGAACGGATATCTTCGGACTGAAGTTCTT GCGACTGAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGGGGGATAACAGTTAGAAATGACTG CTAATACCGCATAAGCGCACAGCTTCGCATGGAGCAGTGTGAAAAACTCCGGTGGTATGAGATGGACCCGCGTCAG ATTAGCTAGTTGGCAGGGTAACGGCCTACCAAGGCGACGATCTGTAGCCGACCTGAGAGGGTGACCGGCCACATTG GGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAG CGACGCCGCGTGAGCGAAGAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGGAAGATAATGACGGTACCTGACTA AGAAGCTCCGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGGAGCAAGCGTTATCCGGATTTACTGGGTGT AAAGGGAGCGTAGACGGTTTGACAAGTCTGATGTGAAATTCCAGGGCTTAACCCTGGACCTGCATTGGAAACTGTC GGACTAGAGTGTCGGAGAGGTGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGT GGCGAAGGCGGCTCACTGGACGATAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTG GTAGTCCACGCCGTAAACGATGTGTACTAGGTGTTGGGGAGCAAAGCTCTTCGGTGCCGTCGCAAACGCAGTAAGT ACACCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATG TGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAATCTTGACATCGGAGTGACCGCTCTTTAATCGGAGCTTTC CTTCGGGACACTCCAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAAC GAGCGCAACCCTTATCCTTAGTAGCCAGCAAGTGAAGTTGGGCACTCTAGGGAGACTGCCAGGGATAACCTGGAGG AAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGATTTGGGCTACACACGTGCTACAATGGCGTAAACAAAGG GAAGCGATCACGTGAGTGTGAGCAAATCTCAAAAATAACGTCCCAGTTCGGACTGTAGTCTGCAACCCGACTACAC GAAGCTGGAATCGCTAGTAATCGCAGGTCAGCATACTGCGGTGAATACGTTCCCGGGTCTTGCACACACCGCCCGT CA GQ448468.1.1366 AGAGTTTGATCCTGGCTCAGGATGAACGCTGACAGAATGCTTAACACATGCAAGTATACTTGATCCTTCGGGTGAT GGTGGCGGACGGGTGAGTAACGCGTAAAGAACTTGCCCTGCAGTCTGGGACAACATTTGGAAACGAATGCTAATCC CGCATAAGCCCACAGCTCGGCATCGAGCAGAGGGAAAAGGAGTGATCTGCTTTGAGATGGCCTCGCGTCCGATTAG CTGGTTGGTGAGGTGACGGCCCATCAAGGCAACGATCGGTAGCCGGACTGAGAGGTTGAACGGCCACATTGGGATT GAGACACGGCCCTTACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGACCAAAAGTCTGATCCAGCAAT TCTGTGTGCACGATGAAGTTTTTCGGAATGTAAAGTGCTTTCAGTTGGGACGAAGTAAGTGACGGTACCAACAGAA GAAGCGACGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTA AAGCGCGTCTAGGCGGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCAA ACTAGAGTACTGGAGAGGTGGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATGG GGAAGCCAGCCCACTGGACAGATACTGACGCTAAAGCGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGT AGTCCACGCCGTAAACGATGATTACTAGGTGTTGGGGGTCGAACCTCAGCGCCCAAGCTAACGCGATAAGTAATCC GCCTGGGGAGTACGTACGCAAGTATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTT TAATTCGACGCAACGCGAGGAACCTTACCAGCGTTTGACATCCTAAGAAATTAGCAGAGATGCTTTTGTGCCCCTT CGGGGGAACTTAGTGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACG AGCGCAACCCCTTTCGTATGTTGCCATCATTAAGTTGGGCACTCATGCGATACTGCCTGCGATGAGCAGGAGGAAG GTGGGGATGACGTCAAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTACAATGGGTAGTACAGAGAGTC GCAAACCTGCGAGGGGGAGCTAATCTCAGAAAACTATTCTCAGTTCGGATTGTACTCTGCAACTCGAGTACATGAA GTTGGAATCGCTAGTAATCGCAAATCAGCTATGTTGCGGTGAATACGTTCTCGGGTCTTGTACACACCGCCCGT EU774020.1.1361 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAACACATGCAAGTCGAGCGATTCTCTTCGGAGAA GAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCCTGTACACACGGATAACATACCGAAAGGTATGCTAATA CGGGATAATATATAAGAGTCGCATGACTTTTATATCAAAGATTTTTCGGTACAGGATGGACCCGCGTCTGATTAGC TTGTTGGCGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCCGACCTGAGAGGGTGATCGGCCACATTGGAACTG AGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCAACGC CGCGTGAGCGATGAAGGCCTTCGGGTCGTAAAGCTCTGTCCTCAAGGAAGATAATGACGGTACTTGAGGAGGAAGC CCCGGCTAACTACATGCCAGCAGCCGCGGTAATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCG CGTCTAGGTGGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCAAACTAG AGTACTGGAGAGGTAGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATGGGGAAG CCAGCCTACTGGACAGATACTGACGCTAAAGCGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCC ACGCCGTAAACGATGATTACTAGGTGTTGGGGGTCGAACCTCAGCGCCCAAGCTAACGCGATAAGTAATCCGCCTG GGGAGTACGTACGCAAGTATGAAACTCAAAGGAGTTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATT CGACGCAACGCGAGGAACCTTACCAGCGTTTGACATCCTAGGAATGAGAAAGAGATTTCTTAGTGCTCCTTCGGGA GAACCTAGAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGC AACCCCTATTGTATGTTGCCATCATTAAGTTGGGCACTCATGCGATACTGCCTGCGATGAGCAGGAGGAAGGTGGG GATGACGTCAAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTACAATGGGCAGTACAGAGAGAAGCAAT ACCGCGAGGTGGAGCCAAACTTAAAAACCAGTCTCAGTTCGGATTGTAGGCTGAAACTCGCCTACATGAAGCTGGA GTTACTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTACCCGGGTCTTGTACACACCGCCCGTCA GQ491183.1.1360 GATGAACCCTTGCGGCGTGCTTAACACATGCAAGTCGAACGGGAAACATTTTATTGAAGCTTCGGCAGATCTAGCT TGTTTCTAGTGGCGGACGGGTGAGTAACGCGTGGGCAACCTGCCTCACACTGGGGGATAACAGTCAGAAATGGCTG CTAATACCGCATAAGCGCACAGCATCGCATGATGCAGTGTGAAAAACTCCGGTGGTGTGAGATGGACCCGCGTTGG ATTAGCTAGTTGGCAGGGCAGCGGCCTACCAAGGCGACGATCCATAGCCGGCCTGAGAGGGTGAACGGCCACATTG GGACTGAGACACGGCCCAGACTCCCACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAG CGACGCCGCGTGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGACGGTACCTGACTA AGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACTGGGTGT AAAGGGAGCGTAGACGGTGTTGCAAGTCTGATGTGAAAGGCGGGGGCTCAACCCCTGGACTGCATTGGAAACTGTG ATACTCGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGAAGGAATACCAGT GGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAAGATTAAGAAACCTC TGGGTAGTCCACGCCCGTAAACGAAGGAATAAAGGGGTCGGGAGCAGAGCTTTTCGGTGCCGCAGCAAACCCAATA AGTATTCCACCTTGAGAGGACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGGACCCGCACAAGCGGTGGAG CATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCCTCTGACCGCACCTTAACCGGTGC TTTCCTTCGGGACAGAGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCG CAACGAGCGCAACCCTTATCCTTAGTAGCCAGCGGTCCGGCCGGGCACTCTGGGGAGACTGCCAGGGATAACCTGG AGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGATTTGGGCTACACACGTGCTACAATGGCGTAAACAA AGGGAAGCGATCACGTGAGTGCGAGCAAATCTCAAAAATAACGTCCCAGTTCGGACTGTAGTCTGCAACCCGACTA CACGAAGCTGGAATCGCTAGTAATCGCAGGTCAGCATACTGCGGTGAATACGTTCCCGGGTCTTGTAC GQ491426.1.1332 GCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAGCGAAGCACTTGCCATTGACTCTTCGGAAGAT TTGGCATTTGACTGAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACGGGGGAATAACAGTTAGAA ATGGCTGCTAATGCCGCATAACCGCACAGGACCGCATGGACTGGTGTGAAAAACTGAGGTGGTATGAGATGGGCCC GCGTCTGATTAGGTTAGTTGGCGGGGTAACGGCCCACCAAGCCGACGATCAGTAGCCGACCTGAGAGGGACCGGCC ACATTGGGACTGAGACATGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGAGGAAACTCTG ATGCAGCGACGCCGCATGAAGGAAGAAGTATCTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGACGGTACC TGACTAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACT GGGTGTAAAGGGAGCGTAGACGGACGGGCAAGTCTGATGTGAAAGCCCGGGGCTTAACCCCGGGACTGCATTGGAA ACTGTCCATCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAAC ACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGAT ACCCTGGTAGTCCACGCCGTAAACGATCAATAATGGGTGTCGGGTTGCAAAGCAATCCGGTGCCGCAGCAAACGCA GTAAGTATTCCCCCTCGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAAGGGACGGGGATCCGCACAAGCGGCGG AGCATGTGGTTTAATTAGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCTGCCTGACCGTTCCTTAACCGGA ACTATCTTTCGGGACAGGCAAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCCTGTCCTTAGTAGCCAGCAGTCCGGCTGGGCACTCTAGGGAGACTGCCGGGGGTAACCC GGAGGAAGGCGGGGAGGAGGTCAAATCATCATGCCCCCCCTGATTTGGGCTACACACGTGGTACAATGGCGTAAAC AAAGGGAAGCGGAGTGGTGACGCTGAGCAAATCTCAAAAATAACGTCCCACTTCGGACTGCAGTCTGCAACTCGAC TGCACGAAGCTGGAATCGCTAGTAATCGCGAATCAGAATG GQ493039.1.1311 GATGAACGCTGGCGGCGTGCCTAACACATGCAAGTCGAGCGATTTACTTCGGTAAAGAGCGGCGGACGGGTGAGTA ACGCGTGGGTAACCTGCCCTGTACACACGGATAACATACCGAAAGGTATGCTAATACGAGATAATATGCTTTTATC GCATGGTAGAAGTATCAAAGCTTTTGCGGTACAGGATGGACCCGCGTCTGATTAGCTAGTTGGTAAGGTAACGGCT TACCAAGGCAACGATCAGTAGCCGACCTGAGAGGGTGATCGGCCACATTGGAACTGAGACACGGTCCAAACTCCTA CGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCAACGCCGCGTGAGCGATGAAGGCCT TCGGGTCGTAAAGCTCTGTCCTCAAGGAAGATAATGACGGTACTTGAGGAGGAAGCCCCGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGGGGCTAGCGTTATCCGGAATTACTGGGCGTAAAGGGTGCGTAGGTGGTTTCTTAAG TCAGAGGTGAAAGGCTACGGCTCAACCGTAGTAAGCCTTTGAAACTGGGAAACTTGAGTGCAGGAGAGGAGAGTGG AATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTTGCGAAGGCGGCTCTCTGGACTGTAAC TGACACTGAGGCACGAAAGCGTGGGAGCAAACAAGATTAGNTNCCCTGGTAGTCCNCGCCGTNNCCGCCCATAAAG AGCTGTCGGAGGTTACCCCCTTCGGTGGCGCAGGTAACGCAATAAAGAATTCCGCCTGGGAAGGAACGCTTCGCAA GAGTGAAATTAAAAGGAATAGACGGGGACCCGCTCAAGTAGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGA ACTTTCTCTAAGCTTGACATCCTTTTGACCGATGCCTAATAGCATCAATCCCTTCTGGGACAGAAGTGACAGGTGG TGCATGGTTGTTGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGCCTTTAGTTG CCAGCATTAAGTTGGGCACTCTATAGGGACTGCCAGGGATAACCTGGAGGAAGGTGGGGATGACGTCAAATCATCA TGCCCCTTATGCTTAGGGCTACACACGTGCTACAATGGGTGGTACAGAGGGCAGCCAAGTCGTGAGGCGGAGCTAA TCCCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGAAACTCGCCTACATGAAGCTGGAGTTACTAGTAATCGCAG ATCAGAATGATGCGGTGAA JN387556.1.1324 CGTAAGTAACCTGCCCTGTACACACGGATAACATACCGAAAGGTATGCTAATACGGGATAATATATTTTGATCGCA TGGTCGAGATATCAAAGCTCCGGCGGTACACCAGGGACCCCCGACAGAGGAGCTAGTTGGTAGTAATGTCACCAAG GCGACGATCAGAAGCCGAACTGAGAGGGGGATCCGCACATGACTGAGACACGGTCAAACTCCTACGGGAGGCAGCA GTGGGGAATATGCCAATGGGCGAAAGCTGATGCAGCACGCGCGTGAGCGATGAGGCTCGGGTCGTAAAGCTCGTCT CAAGGAAGATAATGACGGTACTTGAGGAGGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGG GCTAGCGTTATCCGGAATTACTGGGCGTAAAGGGTGCGTAGGCGGTCTTTCAAGTCAGGAGTGAAAGGCTACGGCT CAACCGTAGTAAGCTCTTGAAACTGTAAGACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAA TGCGTAGATATTAGGAGGAACACCAGTTGCGAAGGCGGCTCTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGT GGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTACTAGCTGTCGGAGGTTACCCCCT TCGGTGGCGCAGCTAACGCATTAAGTACTCCGCCTGGGAAGTACGCTCGCAAGAGTGAAACTCAAAGGAATTGACG GGGACCCGCACAAGTAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTAAGCTTGACATCCCA CTGACCCTTCCCTAATCGGAAGCTTCCCTTCGGGACAGTGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTC GTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGCCTTTAGTTGCCAGCATTAAGTTGGGCACTCTAGA GGGACTGCCAGGGATAACCCGGAGGAGTGGGGATGACGTCAAATCATCATGCCCTTATGCTAGGCTACACACGTGC TACAATGGGTGGTCAGAGGCCAGCCAGTCGTGAGGCCGAGCTATCCCATAAGCCATTCTCGTCCGGATTGTAGGCT GAACTCGCCTACATGAGCTGGAATTACAAGTATGCGATCGATGCTGCGTGATGCGTCCGGGTCTTGTACACACCGC CCGTCACACCATGGGAGTTGGGGGCGCCCGAAGCCGGATTGCTAACCTTTTGGAAGCGTCCGTCGAAGGTGAAACC AATAACTGGGGTGAAGTCGTAACAAGGTAACC EU775983.1.1288 GAAAGCGGCGGACGGGTGAGTAACGCGTAGGCAACCTGCCCCATACAGAGGGATAGCATCTGGAAACGGATATTAA TACCTCATAATACTTAGAGATCACATGGTAACTAAGTCAAAGATTTATCGGTATGGGATGGGCCTGCGTCTGATTA GCTAGTTGGTGGGGTAACGGCTCACCAAGGCGACGATCAGTAGCCGACCTGAGAGGGTGATCGGCCACATTGGAAC TGAGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCAAC GCCGCGTGAGCGATGAAGGCCTTCGGGTCGTAAAGCTCTGTCCTCAAGGAAGATAATGACGGTACTTGAGGAGGAA GCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCTAGCGTTATCCGGATTTACTGGGCGTAAAG GGTGCGTAGGCGGTCTTTCAAGTCAGGAGTTAAAGGCTACGGCTCAACCGTAGTAAGCTCCTGATACTGTCTGACT TGAGTGCAGGAGAGGAAAGCGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTAGCGA AGGCGGCTTTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGT CCACGCTGTAAACGATGAGTACTAGGTGTCGGAGGTTACCCCCTTCGGTGCCGCAGCTAACGCATTAAGTACTCCG CCTGGGGAGTACGCACGCAAGTGTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGTAGCGGAGCATGTGGTTT AATTCGAAGCAACGCGAAGAACCTTACCTAGGCTTGACATCCTTCTGACCGAGGACTAATCTCCTCTTTCCCTCCG GGGACAGAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGC GCAACCCTTGTCTTTAGTTGCCATCATTAAGTTGGGCACTCTAGAGAGACTGCCAGGGATAACCTGGAGGAAGGTG GGGATGACGTCAAATCATCATGCCCCTTATGCCTAGGGCTACACACGTGCTACAATGGGTGGTACAGAGGGCAGCC AAGCCGTGAGGTGGAGCAAATCCCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGAAACTCGCCTACATGAAGCT GGAGTTACTAGTAATCGCAGATCAGAATGCTGCGGTGAATGCGTTCCCGGGTCTTGCACACACCGCCCGTCA OTUs in Table 5 GQ449137.1.1391 CTGGCTCAGGATGAACGCTAGCGACAGGCTTAACACATGCAAGTCGAGGGGCATCACGGGAGGTAGCAATACCTTC TGGTGGCGACCGGCGCACGGGTGAGTAACACGTATGCAACCTGCCCTGTACAGAGGGACAAGCGGTGGAAACGCCG TCTAATCCCGCATGCACTCTTCCGGGGGCATCCCCGGGAGAGTAAAGGAGAGATCCGGTACAGGATGGACATGCGG CGCATTAGTTAGTTGGCGGGGTAACGGCCCACCAAGACGACGATGCGTAGGGGTTCTGAGAGGAAGGTCCCCCACA TTGGAACTGAGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGGCGGAAGCCTGAAC CAGCCAAGTCGCGTGAGGGAAGACGGCCCTACGGGTTGTAAACCTCTTTTGTCGGGGAGCAATGCCGCCTTTGCGA AGGCGGAGGGAGAGTACCCGAAGAAAAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAA GCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTCTGTAAGACAGATGTGAAATCCCCGGGCTCAAC CTGGGAATTGCATTTGTGACTGCAGGACTAGAGTTCATCAGAGGGGGGTGGAATTCCAAGTGTAGCAGTGAAATGC GTAGATATTTGGAAGAACACCAATGGCGAAGGCAGCCCCCTGGGATGCGACTGACGCTCATGCACGAAAGCGTGGG GAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCTACTGGTTGTTGGGGATTAATATCCTTG GTAACGAAGCTAACGCGTGAAGTAGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGG ACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGATACGCGAGGAACCTTACCCGGGCTCAAACGGCACAGT GATACTTTTGAAAGGAGGTAGCTCTACGGAGACTGTGCCGAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAG GTGTCGGCTTAAGTGCCATAACGAGCGCAACCCCTATTGTCAGTTGCCAGCAGGTAAAGCTGGGGACTCTGACGAG ACTGCCGGCGCAAGCTGAGAGGAAGGCGGGGATGACGTCAAATCAGCACGGCCCTTACGTCCGGGGCGACACACGT GTTACAATGGCAGGCACAGCGGGAAGCCACCCGGCGACGGGGAGCGGAACCCGAAAGCCTGTCTCAGTTCGGATCG GAGTCTGCAACTCGACTCCGTGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCC GGGCCTTGTACACACCGCCCGTA HK555938.1.1357 ACGGCACCCCTCTCCGGAGGGAAGCGAGTGGCGAACGGCTGAGTAACACGTGGAGAACCTGCCCCCTCCCCCGGGA TAGCCGCCCGAAAGGACGGGTAATACCGGATACCCCCGGGCGCCGCATGGCGCCCGGGCTAAAGCCCCGACGGGAG GGGATGGCTCCGCGGCCCATCAGGTAGACGGCGGGGTGACGGCCCACCGTGCCGACAACGGGTAGCCGGGTTGAGA GACCGACCGGCCAGATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTGCGCAATG GGGGGAACCCTGACGCAGCGACGCCGCGTGCGGGACGGAGGCCTTCGGGTCGTAAACCGCTTTCAGCAGGGAAGAG TCAAGACTGTACCTGCAGAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCGAGCGTT ATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGGCAGGCCGGGGGTCGAAGCGGGGGGCTCAACCCCCC GAAGCCCCCGGAACCTCCGCGGCTTGGGTCCGGTAGGGGAGGGTGGAACACCCGGTGTAGCGGTGGAATGCGCAGA TATCGGGTGGAACACCGGTGGCGAAGGCGGCCCTCTGGGCCGAGACCGACGCTGAGGCGCGAAAGCTGGGGGAGCG AACAGGATTAGATACCCTGGTAGTCCCAGCCGTAAACGATGGACGCTGGGTGTGGGGGGACGATCCCCCCGTGCCG CAGCCNACGCATTAAGCGTCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGC ACAAGCAGCGGAGCATGTGGCTTAATTCGAAGCAACGCGAAGAACCTTACGGCGCATCCCCCCGAGGCCCACGGGG GGTCCGCCGCGTGGGTCAGAGGAGCGCATACGGGAGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGT TAAGTCCCGCAACGAGCGCAACCCCCGCCGCGTGTTGCCATCGGGTGATGCCGGGAACCCACGCGGGACCGCCGCC GTCAAGGCGGAGGAGGGCGGGGACGACGTCAAGTCATCATGCCCCTTATGCCCTGGGCTGCACACGTGCTACAATG GCCGGTACAGAGGGATGCCACCCCGCGAGGGGGAGCGGATCCCGGAAAGCCGGCCCCAGTTCGGATTGGGGGCTGC AACCCGCCCCCATGAAGTCGGAGTTGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATGCGTTCCCGGGCCTTGT ACACACCGCCCGTCACACCACCCGAGTCGTCTGCACCCGAAGTCGCCGGCCCAACCGCAAGGGGG GQ358246.1.1466 AGAGTTGATCTGGCTCAGATTGAACGCTGGCGGCAGGCTTAATACATGCAAGTCGAACGGTAACAGCAAAAAAGCT TGCTTTTTTGGCTGACGAGTGGCGGACGGGTGAGTAATACCTAGGAAGCTGCCTAAACGAGGGGGATAACACCTGG AAACGGGTGCTAATACCGCATGATACCGCAAGGTCAAAGGTTGGTTTACCAATCGCGTTTAGATGCGCCTAGGAGG GATTAGCTAGTTGGTGGGGTAACGGCTCACCAAGGCGATGATCAGTAGCCGGTCTGAGAGGATGAACGGCCACATT GGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTTCCGCAATGGGCGAAAGCCTGACGGA GCAATGCCGCGTGAGTGATGAAGGGATTCGTCCCGTAAAGCTCTGTTGTATATGACGAATGTGCAGATTGTGAATA ATGATTTGTAATGACGGTAGTATACGAGGAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGG CGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCATGTAGGCGGTTTTTTAAGTCTGGAGTGAAAATGCGGGGCTC AACCCCGTATGGCTCTGGATACTGGAAGACTTGAGTGCAGGAGAGGAAAGGGGAATTCCCAGTGTAGCGGTGAAAT GCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGCCTTTCTGGACTGTGTCTGACGCTGAGATGCGAAAGCCAG GGTAGCGAACGGGATTAGATACCCCGGTAGTCCTGGCCGTAAACGATGGGTACTAGGTGTGGGAGGTATCGACCCC TTCCGTGCCGGAGTTAACGCAATAAGTACCCCGCCTGGGGAGTACGTCCGCAAGGATGAAACTCAAAGGAATTGAC GGGGGCCCGCACAAGCGGTGGAGTATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCAAGGCTTGACATTGA TTGAAAGACCTAGAGATAGGTCCCTCTCTTCGGAGACAAGAAAACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTC GTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCCTATGTTACCAGCGGGTAATGCCGGGGACTCAT AGGAGACTGCCAAGGACAACTTGGAGGAAGGCGGGGATGACGTCAAGTCATCATGCCCCTTATGTCTTGGGCTACA CACGTACTACAATGGTCGGCAACAGAGGGAAGCAAAGCCGCGAGGCAGAGCAAACCCCAGAAACCCGATCTCAGTT CGGATTGCAGGCTGCAACTCGCCTGCATGAAGTCGGAATCGCTAGTAATCGCAGGTCAGCATACTGCGGTGATTAC TATCCCGGGCGTTGTACTCACCGCCCGTCAGGCGGAGTTCGTACTTCAAATGTGCCACACTGGG New.ReferenceOTU82 TACGTAGGTGGCAAGCGTTGTCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATTTTTAAGTGGGATGTGAAA TACCCGGGCTCAACCTGGGTGCTGCATTCCAAACTGGAAATCTAGAGTGCAGGAGGGGAAAGTGGAATTCCTAGTG TAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGACTTTCTGGACTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGG New.ReferenceOTU52 TACGTAGGTGGCGAGCGTTATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATGATTAAGTGGGATGTGAAA TACCCGGGCTCAACTTGGGTGCTGCATTCCAAACTGGTTATCTAGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTG TAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGACTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGG GQ138615.1.1402 AGAGTTTGATCCTGGCTCAGGATGAACGCTAGCGATAGGCCTAACACATGCAAGTCGAGGGGCAGCACATGAGTAG CAATACGATGGTGGCGACCGGCGCACGGGTGAGTAACACGTATGCAACCTACCTTTAACAGGGGAATAACCCGTTG AAAAACGGACTAATACTCCATAACACAGGGGTCCCGCATGGGAATATTTGTTAAAGATTTATCGGTTGAAGATGGG CATGCGTTCCATTAGCTAGTTGGTAGGGTAAAGGCCTACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGAAC GGCCACACTGGGACTGAGACACGGCCCACACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGCGAAAG CCTGACCGAGCAACGCCGCGTGAATGATGAAGGCCTTCGGGTTGTAAAATTCTGTTATAAGGGAAGAACGACTTTA GTAGGAAATGGCTAGAGTGTGACGGTACCTTATGAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATAC GTAGGTGGCGAGCGTTATCCGGAATTATTGGGCGTAAAGAGCGCGCAGGTGGTTGATTAAGTCTGATGTGAAAGCC CACGGCTTAACCGTGGAGGGTCATTGGAAACTGGTCGACTTGAGTGCAGAAGAGGGAAGTGGAATTCCATGTGTAG CGGTGAAATGCGTAGAGATATGGAGGAACACCAGTGGCGAAGGCGGCTTCCTGGTCTGTAACTGACACTGAGGCGC GAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTGGGGGT CGAACCTCAGTGCTGAAGTTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAA TTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGAC ATACCATTGACCGTTCTAGAGATAGGATTTTCCCTTCGGGGACAATGGATACAGGTGGTGCATGGTTGTCGTCAGC TCGTGTCGTGAGATGTTGGGTTAGGTCCCGCAACGAGCGCAACCCCTGTCGTTAGTTGCCAGCATTCAGTTGGGGA CTCTAACGAGACTGCCAGTGACAAACTGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGG CTACACACGTGCTACAATGGTTGGTACAAAGAGAAGCGAAGCGGTGACGTGGAGCAAACCTCATAAAGCCAATCTC AGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTTGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGA ATACGTTCCCGGGTCTTGTACACACCGCCCGTCA JN681884.1.1409 TGCAAGTAGAACGCTGAAGACTGGTGCTTGCACCGGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACC TGCCTGATAGCGGGGGATAACTATTGGAAACGATAGCTAATACCGCATAACAGGGAATAACACATGTTATTTTTTT GAAAGGGGCAATTGCTCCACTATCAGATGGACCTGCGTTGTATTAGCTAGTAGGTGAGGTAACGGCTCACCTAGGC GACGATACATAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCA GCAGTAGGGAATCTTCGGCAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGT AAAGCTCTGTTGTAAGAGAAGAACGTTGAGTAGAGTGGAAAGTTACTCAAGTGACGGTATCTTACCAGAAAGGGAC GGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGC GCAGGCGGTTTAATAAGTCTGAAGTTAAAGGCAGTGGCTCAACCATTGTTCGCTTTGGAAACTGTTAAACTTGAGT GCAGAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCG GCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCACG CCGTAAACGATGAGTGCTAGGTGTTGGGTCCTTTCCGGGACTCAGTGCCGACGCTAACGCATTAAGCACTCCGCCT GGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAAT TCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCGATGCTATCCCTAGAGATAGGGAGTTACTTCGGTAC ATCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAAC CCCTATTGTTAGTTGCCATCATTCAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGAT GACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTACAACGAGTTGCGAGTCG GTGACGGCAAGCTAATCTCTTAAAGCCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAAT CGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAG AGTTTGTAACACCCAAAGTCGGTGAGGTAACCTTCGGAGCC GU303759.1.1517 AGAGTTTGATCATGGCTCAGGACGAACGCCGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGACTTTAGCT TGCTAAAGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGATAACTATTGGA AACGATAGCTAATACCGTATAACAGCATTTAACACATGTTAGATGCTTGAAAGGAGCAATTGCTTCACTAGTAGAT GGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGACCTGAGAGGGTG ATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGC AACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGT GTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTA ATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAA AGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTG TAGCGGTGAAATGCGTAGATATATGGARGGAAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGA GGCTCGAGAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAGCGATGAGTGCTAGGTGTT AGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAA CTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACC AGGTCTTGACATCCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACATCGGTGACAGGTGGTGCATGGTTG TCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCATTAA GTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTAT GACCTGGGCTACACACGTGCTACAATGGCGGTCAACAGAGGGAAGCAATACTGTGAAGTGGAGCAAACCCCTAAAA GCCGTCCCAGTTCGGATTGCAGGCTGCAACCCGCCTGTATGAAGTTGGAATCGCTAGTAATCGCGGATCAGCATGC CGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTCGGGAACACCCGAAGTCCGTAG CCTAACTTTCACGAGGGGGCGCGGCCGAAGGTGGGTTCGATAATTGGGGTGAAGTCGTAACAAGGTAACCGTA New.ReferenceOTU114 TACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAAA GGCAGTGGCTTAACCATTTTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTGT AGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGGC TCGAAAGCGTGGGGAGCAAACAGG EU774881.1.1422 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAGCGGAACTAACAGA TTTACTTCGGTAATGACGTTAGGAAAGCGAGCGGCGGATGGGTGAGTAACACGTGGGGAACCTGCCCCATAGTCTG GGATACCACTTGGAAACAGGTGCTAATACCGGATAAGAAAGCAGATCGCATGATCAGCTTTTAAAAGGCGGCGTAA GCTGTCGCTATGGGATGGCCCCGCGGTGCATTAGCTAGTTGGTAAGGTAAAGGCTTACCAAGGCAATGATGCATAG CCGAGTTGAGAGACTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAAT CTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTG TTGGTGAAGAAGGATAGAGGTAGTAACTGGCCTTTATTTGACGGTAATCAACCAGAAAGTCACGGCTAACTACGTG CCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAA TAAGTCTGAAGTTAAAGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGA GTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTG TAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGA GTGCTAGGTGTTAGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACC GCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCG AAGAACCTTACCAGGTCTTGACATCCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACATCGGTGACAGGT GGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGT TGCCATCATTAAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAGTCAT CATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGTACAACGAGAAGCGAGCCTGCGAAGGCAAGCG AATCTCTGAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACTCGACTGCACGAAGCTGGAATCGCTAGTAATCGC GGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGCACACACCGCCCGTCA AB469559.1.1551 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTGGAACGCACAGTTAGTATGTA GTTTACTACAACATTACTTGTGAGTCGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTTGTAGCGGGGGATAAC TATTGGAAACGATAGCTAATACCGCATAACAGTTGATAACTCATGTTATTAGCTTGAAAGATGCAACAGCATCACT ACGAGATGGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAAAGGCTCACCAAGGCCACGATACATAGCCGACCTGA GAGGGTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAA TGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAG AACGTTGATGAGAGTGGAAAATTCATCAAGTGACGGTATCTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGC CGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTCGTAAGTCTG AAGTTAAAGGCAGTGGCTCAACCATTGTTCGCTTTGGAAACTGCGAGACTTGAGTGCAGAAGGGGAGAGTGGAATT CCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGAC GCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGG TGTTAGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTT GAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCT TACCAGGTCTTGACATCCCGATGCCCGCTCTAGAGATAGAGTTTTACTTTTGTACATCGGTGACAGGTGGTGCATG GTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCA TTGAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCC TTATGACCTGGGCTACACACGTGCTACAATGGCTGGTACAACGAGTCGCAAGCCGGTGACGGCAAGCTAATCTCTT AAAGCCAGTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGC ACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCG GTGAGGTAACCTTTTAGGAGCCAGCCGCCTAAGGTGGGATAGATGATTGGGGTGAAGTCGTAACAAGGTAGCCGTA TCGGAAGGTGCGGCTG HK557089.3.1395 AGACTTTAGCTTGCTAAAGTTGGAAGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTACTAGCGGGGGA TAACTATTGGAAACGATAGCTAATACCGCATAACAGCATTTAACCCATGTTAGATGCTTGAAAGGAGCAATTGCTT CACTAGTAGATGGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATACATAGCCGAC CTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCG GCAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGA GAAGAACGTGTGTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAG TCTGAAGTTAAAGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAGACTTGAGTGCAGAAGGGGAGAGTGG AATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAAC TGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGC TAGGTGTTAGGCCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAA GGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGA ACCTTACCAGGTCTTGACATCCCGATGCTATTCCTAGAGATAGGAAGTTTCTTCGGAACTGTGAGACTTGAGGGCA GAAGGGTAGAGTGCACTTGTATGGGGAGCTGTGGAATGCGTTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCC ATCATTAAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATG CCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTACAACGAGTCGCGAGTCGGTGACGGCAAGCAAATC TCTTAAAGCCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGAT CAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAA GTCGGTGAGGTANCCTTTTAGGAGC EU358719.1.1513 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTAGAACGCTGAAGAAAGGAGCT TGCTTCTTTTGGATGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTTGTAGCGGGGGATAACTATTGGA AACGATAGCTAATACCGCATAACAGCTTTTGACACATGTTAGAAGCTTGAAAGATGCAATTGCATCACTACGAGAT GGACCTGCGTTGTATTAGCTAGTAGGTAGGGTAACGGCCTACCTAGGCGACGATACATAGCCGACCTGAGAGGGTG ATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGC AACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGT GTGAGAGTGGAAAGTTCACACAGTGACGGTAACTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTA ATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAA AGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAAACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTG TAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGG CTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTAGG CCCTTTCCGGGGCTTAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTC AAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGG TCTTGACATCCCAGTGACCGCTCTAGAGATAGAGTTTTTCTTCGGAACACTGGTGACAGGTGGTGCATGGTTGTCG TCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTGTTAGTTGCCATCATTCAGTT GGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGAC CTGGGCTACACACGTGCTACAATGGTTGGTACAACGAGTCGCAAGTCGGTGACGGCAAGCAAATCTCTTAAAGCCA ATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGCAGGAATTGCTAGTAATGGCAGGTCAGCATACTGC CGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTCTGTAACACCCGAAGTCGGTAGTCT AACTACGGAGGACGCCGCCGAAGGTGGGACAGATAATTGGGGTGAAGTCGTAACAAGGTAGCCGTA HQ748204.1.1442 CTAATACATGCGAGGAGAACGCTGAAGACTTTCTTTTGCTATAGTTGGGAGAGTTGCTAACGGGTGAGTAACGCGT AGGTGACCTGCCTACTAGCGGGGGATAACTATTGCAAACGATAGCTAATACCGCATAACAGCCTTTAACCCATGTT AGATGCTTGAAAGGAGCAATTGCTTCACTAGTAGATGGACCTGCGTTGTATTAGCTAGTTGGTGAGGTAACGGCTC ACCAAGGCGACGATACATAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCATACTCCTAC GGGAGGCACCAGTAGGGAATCTTCGGGAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTT CGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGTGTGTGAGAGTGGAAAGTTCACACTGTGACGGTAACTTACCAG AAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTA AAGCGAGCGCAGGCGGTTTAATAAGTCTGAAGTTAAAGGCAGTGGCTTAACCATTGTTCGCTTTGGAAACTGTTAG ACTTGAGTGCATAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGG CGAAAGCGGCTCTCTGGTCTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGT AGTCCACGCTGTAAACGATGAGTGGTAGGTGTTAGGCCCTTTCTGGGGTTTAGTGCCGCAGATTACGCATTAAGCC ATTCGCCTGGGGAGTACGACCGCAAGGTTGAAACTTAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGT GGTTTAATTAGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCGATGCTATTCTTAGAGATAGGAAGTTTC TTCGGAACATCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGAGAGATGTTGGGTTAAGTCCCTCAACG AGCGCAACCCCTATTTTTATTTGCCATCATTAAGTTGGGCAATCTAGCGAGACTGCCGGTAATAAACCGGAGGAAG GTGGGGATGACGTCAAATCATCATGCTCCTTATGTCATGGGGTACACACGTGGTACAATGGTTGGTACAACGAGTC GCGAGTTGGTGAAGGCAAGCAAATCTCTTAAAGCCAATATCAGTTCGGATTGTAGGCTGCAAATAGCCTACATGTA GTCGGAATTGTTAGTAATCGGGGATCAGCACTCCGCGGTGAATACGTTTCCGGGCCTTGTACACCCCGCCCGTCTA CACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACTCTTTTAGGAGCCAGCCGCCTAAGGTGGGATAGA GQ338727.1.1397 CTACCTGCAGTCGACGAACACCTTATTTGATTTTCTTCGGAACTGAAGATTTGGTGATTGAGTGGCGGACGGGTGA GTAACGCGTGGGTAACCTGCCCTGTACAGGGGGATAACAGTCAGAAATGACTGCTAATACCGCATAAGACCACAGC ACCGCATGGTGCAGGGGTAAAAACTCCGGTGGTACAGGATGGACCCGCGTCTGATTAGCTGGTTGGTGAGGTAACG GCTCACCAAGGCGACGATCAGTAGCCGGCTTGAGAAAGTGAACGGCCACATTGGGACTGAGACACGGCCCAAACTC CTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCGACGCCGCGTGAGTGAAGAAGT ATCTCGGTATGTAAAGCTCTATCAGCAGGGAAGAAAATGACGGTACCTGACTAAGAAGCCCCGGCTAACTACGTGC CAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGTGGTATGGC AAGTCAGAAGTGAAAACCCAGGGCTTAACTCTGGGACTGCTTTTGAAACTGTCAGACTGGAGTGCAGGAGAGGTAA GCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCTTACTGGACTG AAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGA ATACTAGGTGTCGGGGCCGTAGAGGCTTCGGTGCCGCAGCCAACGCAGTAAGTATTCCACCTGGGGAGTACGTTCG CAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGTAGCGGAGCATGTGGTTTAATTCGAAGCAACGCGA AGAACCTTACCTAAGCTTGACATCCTTTTGACCGATGCCTAATCGCATCTTTCCCTTCGGGGACAGAAGTGACAGG TGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGCCTTTAG TTGCCATCATTAAGTTGGGCACTCTAGAGGGACTGCCAGGGATAACCTGGAGGAAGGTGGGGATGACGTCAAATCA TCATGCCCCTTATGCTTAGGGCTACACACGTGCTACAATGGGTGGTACAGAGGGCAGCGAAGTCGTGAGGCCAAGC TAATCCCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGAAACCCGCCTACATGAAGCTGGAGTTACTAGTAATCG CAGATCAGAATGCTGCGGTGAATGCGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTTGGGGGCGC CCGAAGCCGGCTAGCTACTTTGGAAGCGT HQ803964.1.1435 GGGGGGCTTAACACATGCAAGTCGAACGAAGCGCTTTCGCTTTAATCTTCGGAGGAAAGAGGAAGTGACTGAGTGG CGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGGGGGATAACAGTTAGAAATGGCTGCTAATACCGCAT AAGCATACAGCACCGCATGGTGCAGTGTGAAAAACTCCGGTGGTATAAGATGGACCCGCGTCTGATTAGGTAGTTG GTGGGGTAACGGCCTACCAAGCCGACGATCAGTAGCCGACCTGAGAGGGTGACCGGCCACATTGGGACTGAGACAC GGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCGACGCCGCGTG AAGGAAGAAGTATTTCGGTATGTAAACTTCTATCAGCAGGGAAGAAAATGACGGTACCTGACTAAGAAGCCCCGGC TAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTA GACGGAAGTGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGTGACTGCTTTGGAAACTGTGCTTCTAGAGTGT CGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGC TTACTGGGCGATAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCC GTAAACGATGAATACTAGGTGTCGGGAAGCACAGCTTTTCGGTGCCGCCGCAAACGCATTAAGTATTCCACCTGGG GAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCG AAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCCGGTGACCGGACAGTAATGTGTCCTTTTCTTCGGAACACC GGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCT TATCCCCAGTAGCCAGCGGTTCGGCCGGGCACTCTGAGGAGACTGCCAGGGATAACCTGGAGGAAGGTGGGGATGA CGTCAAATCATCATGCCCCTTATGACTTGGGCTACACACGTGCTACAATGGCGTAAACAAAGGGAAGCGAGACCGT GAGGTGGAGCAAATCCCAAAAATAACGTCTCAGTTCGGACTGTAGTCTGCAACCCGACTACACGAAGCTGGAATCG CTAGTAATCGCAGATCAGAATGCTGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAG TTGGAAATGCCCGAAGTCAGTGACCCAACCGCAAGGAGGGAGCTGCCGAAGGCAGGTTCGATAACTG FJ951866.1.1493 AGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAAGCACTTTAACTTG ATTTTTTCGGAATGATTGTTCTTGTGACTGAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGG GGGATAACAGTTAGAAATGACTGCTAATACCGCATAAGCGCACGGTATCGCATGATACAGTGTGAAAAACTCCGGT GGTATGAGATGGACCCGCGTCTGATTAGCTAGTTGGCGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCCGACC TGAGAGGGTGACCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCA CAATGGGCGAAAGCCTGATGCAGCGACGCCGCGTGAACGAAAAAGTATTTCGGTATGTAAAGTTCTATCAGCAGGG AAGATAATGACGGTACCTGACTAAGAAGCACCGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGGTGCAAG CGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGTACGGCAAGTCTGATGTGAAAGCCCGGGGCTCAACC CCGGTACTGCATTGGAAACTGTCGAACTAGAGTGTCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCG TAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGG AGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGAATACTAGGTGTGGGAGGACTGACCCCTTCC GTGCCGCAGTTAACACAATAAGTATTCCACCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGG GCCCGCACAAGCAGTGGATTATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCAGGACTTGACATCCAACTA ACGAAGTAGAGATACATTAGGTGCCCTTCGGGGAAAGTTGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCG TGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCTACGCAAGAGCACTCTAGCGAGACT GCCGTTGACAAAACGGAGGAAGGCGGGGACGACGTCAAATCATCATGCCCCTTATGTCCTGGGCTACACACGTAAT ACAATGGCCGTCAACAAAGGGAAGCAAAGCCGCGAGGTGGAGCAAATCCCCAAAAACGGTCTCAGTTCGGATTGCA GGCTGCAACTCGCCTGCATGAAGCTGGAATTGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCCCGGG CCTTGTACACACCGCCCGTCACACCATGAGAGTCGGGAACACCCGAAGTCCGTAGTCTAACCGCAAGGAGGGCGCG GCCGAAGGTGGGTCCGGTAATTGGGGTGAAGTCGTAACAAGGTAACCGT EU772870.1.1289 AGTGGCGAACGGGTGAGTAACGCGTGAGGAACCTGCCTTTCAGTGGGGGACAACAGTTGGAAACGACTGCTAATAC CGCATGATACTTTTTGGAGGCATCTCTGAAAAGTCAAAGCTTTATGTGCTGAAAGATGGTCTCGCGTCTGATTAGC TAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATCAGTAGCCGGTCTGAGAGGATGAACGGCCACATTGGGACTG AGATACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGGCAATGGGGGAAACCCTGACCCAGCAACGC CGCGTGAAGGAAGAAGGCCTTCGGGTTGTAAACTTCTTTTACCAGGGACGAAGAACGTGACGGTACCTGGAGAAAA AGCAACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTTGCAAGCGTTATCCGGATTTATTGGGCGTAAA GCGCGTCTAGGCGGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCTAAAC TAGAGTACTGGAGAGGTAGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATGGGG AAGCCAGCCTACTGGACAGATACTGACGCTAAAGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAG TCCACGCTGTAAACGATGAGTACTAGGTGTCGGAGGTTACCCCCTTCGGTGCCGCAGCTAACGCATTAAGTACTCC GCCTGGGGAGTACGCACGCAAGTGTGGAACTCAAAGGAATTGACGGGGACCCGCACAAGTAGCGGAGCATGTGGTT TAATTCGAAGCAACGCGAAGAACCTTACCTAGGCTTGACATCCTTCTGACCGAGGACTAATCTCCTCTTTCCCTCC GGGGACAGAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAG CGCAACCCTTGTCTTTAGTTGCCATCATTTAGTTGGGCACTCTGGAGAGACTGCCAGGGATAACCTGGAGGAAGGT GGGGATGACGTCAAATCATCATGCCCCTTATGCCTAGGGCTACACACGTGCTACAATGGGTGGTACAGAGGGCAGC TAAGCCGTGAGGTGGAGCAAATCCCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGAAACTCGCCTACATGAAGC TGGAGTTACTAGTAATCGCAGATCAGAATGCTGCGGTGAATGCGTTCCCGGGTCTTGTACACACCGCCCGTCA GQ448468.1.1366 AGAGTTTGATCCTGGCTCAGGATGAACGCTGACAGAATGCTTAACACATGCAAGTATACTTGATCCTTCGGGTGAT GGTGGCGGACGGGTGAGTAACGCGTAAAGAACTTGCCCTGCAGTCTGGGACAACATTTGGAAACGAATGCTAATCC CGCATAAGCCCACAGCTCGGCATCGAGCAGAGGGAAAAGGAGTGATCTGCTTTGAGATGGCCTCGCGTCCGATTAG CTGGTTGGTGAGGTGACGGCCCATCAAGGCAACGATCGGTAGCCGGACTGAGAGGTTGAACGGCCACATTGGGATT GAGACACGGCCCTTACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGACCAAAAGTCTGATCCAGCAAT TCTGTGTGCACGATGAAGTTTTTCGGAATGTAAAGTGCTTTCAGTTGGGACGAAGTAAGTGACGGTACCAACAGAA GAAGCGACGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTA AAGCGCGTCTAGGCGGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCAA ACTAGAGTACTGGAGAGGTGGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATGG GGAAGCCAGCCCACTGGACAGATACTGACGCTAAAGCGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGT AGTCCACGCCGTAAACGATGATTACTAGGTGTTGGGGGTCGAACCTCAGCGCCCAAGCTAACGCGATAAGTAATCC GCCTGGGGAGTACGTACGCAAGTATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTT TAATTCGACGCAACGCGAGGAACCTTACCAGCGTTTGACATCCTAAGAAATTAGCAGAGATGCTTTTGTGCCCCTT CGGGGGAACTTAGTGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACG AGCGCAACCCCTTTCGTATGTTGCCATCATTAAGTTGGGCACTCATGCGATACTGCCTGCGATGAGCAGGAGGAAG GTGGGGATGACGTCAAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTACAATGGGTAGTACAGAGAGTC GCAAACCTGCGAGGGGGAGCTAATCTCAGAAAACTATTCTCAGTTCGGATTGTACTCTGCAACTCGAGTACATGAA GTTGGAATCGCTAGTAATCGCAAATCAGCTATGTTGCGGTGAATACGTTCTCGGGTCTTGTACACACCGCCCGT EU774020.1.1361 AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAACACATGCAAGTCGAGCGATTCTCTTCGGAGAA GAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCCTGTACACACGGATAACATACCGAAAGGTATGCTAATA CGGGATAATATATAAGAGTCGCATGACTTTTATATCAAAGATTTTTCGGTACAGGATGGACCCGCGTCTGATTAGC TTGTTGGCGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCCGACCTGAGAGGGTGATCGGCCACATTGGAACTG AGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCAACGC CGCGTGAGCGATGAAGGCCTTCGGGTCGTAAAGCTCTGTCCTCAAGGAAGATAATGACGGTACTTGAGGAGGAAGC CCCGGCTAACTACATGCCAGCAGCCGCGGTAATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCG CGTCTAGGTGGTTTGGTAAGTCTGATGTGAAAATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCAAACTAG AGTACTGGAGAGGTAGGCGGAACTACAAGTGTAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATGGGGAAG CCAGCCTACTGGACAGATACTGACGCTAAAGCGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCC ACGCCGTAAACGATGATTACTAGGTGTTGGGGGTCGAACCTCAGCGCCCAAGCTAACGCGATAAGTAATCCGCCTG GGGAGTACGTACGCAAGTATGAAACTCAAAGGAGTTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATT CGACGCAACGCGAGGAACCTTACCAGCGTTTGACATCCTAGGAATGAGAAAGAGATTTCTTAGTGCTCCTTCGGGA GAACCTAGAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGC AACCCCTATTGTATGTTGCCATCATTAAGTTGGGCACTCATGCGATACTGCCTGCGATGAGCAGGAGGAAGGTGGG GATGACGTCAAGTCATCATGCCCCTTATACGCTGGGCTACACACGTGCTACAATGGGCAGTACAGAGAGAAGCAAT ACCGCGAGGTGGAGCCAAACTTAAAAACCAGTCTCAGTTCGGATTGTAGGCTGAAACTCGCCTACATGAAGCTGGA GTTACTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTACCCGGGTCTTGTACACACCGCCCGTCA HQ782658.1.1415 AATGCTTAACACATGCAAGTCTACTTGATCCTTCGGGTGATGGTGGCGGACGGGTGAGTAACGCGTAAAGAACTTG CCTTGCAGTCTGGGACAACGTCTGGAAACGGACGCTAATACCGGATATTATGCGAGAGTCGCATGGCTCTTTCATG AAAGCTATATGCGCTGCAGGAGAGCTTTGCGTCCCATTAGTTAGTTGGTGAGGTAACGGCTCACCAAGACCGCGAT GGGTAGCCGGCCTGAGAGGGTGAACGGCCACAAGGGGACTGAGACACGGCCCTTACTCCTACGGGAGGCAGCAGTG GGGAATATTGGACAATGGACCAAAAGTCTGATCCAGCAATTCTGTGTGCACGATGACGGTCTTAGGATTGTAAAGT GCTTTCAATCGGGAAAAAGAAAGTGATGGTACCGATAGAAGAAGCGACGGCTAAATACGTGCCAGCAGCCGCGGTA ATACGTATGTCGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGCGTCTAGGCGGTCTGGTAAGTCTGATGTGGA AATGCGGGGCTCAACTCCGTATTGCGTTGGAAACTGCCAGACTAGAGTACTGGAGAGGTGGGCGGAACTACAAGTG TAGAGGTGAAATTCGTAGATATTTGTAGGAATGCCGATAGAGAAGTCAGCTCACTGGACAGATACTGACGCTGAAG CGCGAAAGCATGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGATTACTAAGCGTCGGG GGTCGAACCTCGGCACTCAAGCTAACGCGATAAGTAATCCGCCTGGGGAGTACGTACGCAAGTATGAAACTCAAAG GAATTGACGGGGACCCGCACAAGTGGTGGAGCATGTGGTTTAATTGGACGCAACGCGAGGAACTTTACCAGCGTGT GACATCCTAGGAATGAGAAAGAGATTTTTCAGTGCTCCTTCGGGAGAACCCAGAGACAGGTGGTGCATGGCTGTGG TCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTATGTTGCCATCATTAAGTT GGGCAATCATGCGATGCTGCCTGCGACGAGCAGGAGGAAGGTGGGGATGAGGTCAAGTCATCATGCCCGTTATATG CTGGGCTACACACGTGCTACAATGGGCAGTACAGAGAGAAGCAAATATGCGAGGAGGAGCAAATGTCAGAAAGCTG TTCGTAGTTCGGATTGTACTCTGCAACTGGAGTACATGAAGTTGGAATCAGTAGTAATCGCAAATCAGCAATGTTG CGGTGAATACGTTCTCGGGTCTGGTACACACCGCCCGTCACACCACGAGAGTTGATTGCACCTGAAGTAGCAGGCC TAACCGTAAGGAAGGGTGGTCCGAGGGTGTGGTTAGCGATTGGGGTG DQ794633.1.1395 AGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAGCGAAGCGCTTTTACGGA TTTCTTCGGATTGAAGTGATTGTGACTGAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGGGG GATAACAGTTAGAAATGACTGCTAATACCGCATAAGCGCACAGTACCGCATGGGTACGGTGTGAAAAACTCCGGTG GTATGAGATGGACCCGCGTCTGATTAGGTAGTTGGTGGGGTAACGGCCTACCAAGCCAACGATCAGTAGCCGACCT GAGAGGGCGACCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCAC AATGGGGGAAACCCTGATGCAGCGACGCCGCGTGAAGGATGAAGTATTTCGGTATGTAAACTTCTATCAGCAGGGA AGAAAATAACGGTACCTGAGTAAGAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGC GTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGAAGTGCAAGTCTGATGTGAAAACCCGAGGCTCAACCA CGGGACTGCATTGGAAACTGTGCTTCTAGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGT AGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGA GCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGACTTACTAGGGTGTCGGGCAGCAAAGCTGTTC GGTTGCCGCAGCCATCGCAATAAGTAGTCCACCTGGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACG GGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGCTCTTGACATCCCT CTGACCGGCAAGTAATGTTGCCTTTCCTTCGGGACAGAGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCG TGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCTTCAGTAGCCAGCATTTAAGGTGGGCACTCAGGA GAGACTGCCAGGGATAACCTGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGAGCAGGGCTACACA CGTGCTACAATGGCGTAAACAAAGGGAAGCGAAAGGGTGACCTGGAGCAAATCTCAGAAATAACGTCTCAGTTCGG ATTGTAGTCTGCAACTCGACTACATGAAGCTGGAATCGCTAGTAATCGCGAATCAGCATGTCGCGGTGAATACGTT CCCGGGTCTTGTACTCACCGCCCGTCA FN668375.4306350.4307737 AGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGCGTGCCTAACACATGCAAGTTGAGCGATTTACTTCGGTAAA GAGCGGCGGACGGGTGAGTAACGCGTGGGTAACCTACCCTGTACACACGGATAACATACCGAAAGGTATGCTAATA CGGGATAATATATTTGAGAGGCATCTCTTGAATATCAAAGGTGAGCCAGTACAGGATGGACCCGCGTCTGATTAGC TAGTTGGTAAGGTAACGGCTTACCAAGGCGACGATCAGTAGCCGACCTGAGAGGGTGATCGGCCACATTGGAACTG AGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCAACGC CGCGTGAGTGATGAAGGCCTTCGGGTCGTAAAACTCTGTCCTCAAGGAAGATAATGACGGTACTTGAGGAGGAAGC CCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCTAGCGTTATCCGGATTTACTGGGCGTAAAGGG TGCGTAGGCGGTCTTTCAAGTCAGGAGTGAAAGGCTACGGCTCAACCGTAGTAAGCTCTTGAAACTGGGAGACTTG AGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTTGCGAAG GCGGCTCTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCC ACGCTGTAAACGATGAGTACTAGGTGTCGGGGGTTACCCCCTTCGGTGCCGCAGCTAACGCATTAAGTACTCCGCC TGGGAAGTACGCTCGCAAGAGTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGTAGCGGAGCATGTGGTTTAA TTCGAAGCAACGCGAAGAACCTTACCTAAGCTTGACATCCCAATGACATCTCCTTAATCGGAGAGTTCCCTTCGGG GACATTGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGC AACCCTTGTCTTTAGTTGCCATCATTAAGTTGGGCACTCTAGAGAGACTGCCAGGGATAACCTGGAGGAAGGTGGG GATGACGTCAAATCATCATGCCCCTTATGCTTAGGGCTACACACGTGCTGATTATGCTAAGGAAATAGGATTTACT GGACAATTCTTAATAGAGCCTAAGCCAAAAGAGCCTACTAAACATCAATATGATTTTGATACTGCTACTGTTTTAG GATTTTTAAGAAAGTATAATCTGGATAAATACTTCAAAGTGAATATAGAAGCAAACCATGCAACACTTGCAGGACA TACTTTCCAACATGAATTAA GQ867445.1.1457 CGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAAGCGATTTGGAGGAAGTTTTCGGATGAAATCTGAATTGAC TGAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCACACAGGGGGACAACAGTTAGAAATGGCTGCTAAT ACCGCATAAGCGCACAGCTTCGCATGAAGCAGTGTGAAAAACTCCGGTGGTGTGAGATGGACCCGCGTCTGATTAG GTAGTTGGTGGGGTAACGGCCTACCAAGCCGACGATCAGTAGCCGACCTGAGAGGGTGACCGGCCACATTGGGACT GAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCGACG CCGCGTGAGTGAAGAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGGAAGAAAATGACGGTACCTGACTAAGAAG CCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGATTTACTGGGTGTAAAGG GAGCGTAGACGGCTTGGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGTCAGGCT AGAGTGCTGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCATAGATATTAGGAGGAACACCAGTGGCGA AGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGT CCACGCCGTAAACGATGAATACTAGGTGTTGGGGAGCAAAGCTCTTCGGTGCCGTCGCAAACGCAATAAGTATTCC ACCTGGGAAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTT TAATTCGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCCATTGAAAAGCCCGTAACGGGGTTCCCTCTTCG GAGCAATGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCG CAACCCTTATCCTAAGTAGCCAGCAGGTAGAGCTGGGCACTCTTGGGAGACTGCCAGGGACAACCTGGAGGAAGGT GGGGATGACGTCAAATCATCATGCCCCTTATGATTTGGGCTACACACGTGCTACAATGGCGTAAACAAAGGGAAGC GAAGCTGTGAAGCTAAGCAAATCTCAAAAATAACGTCTCAGTTCGGATTGTAGTCTGCAACTCGACTACATGAAGC TGGAATCGCTAGTAATCGCGGATCAGAATGCCGCGGTGAACACGTTCCCGGGTCTTGTACACACCGCCCGTCACAC CATGGGAGTCAGTAACGCCCGAAGCCAGTGACCTAACCGCAAGGAAGGAGCTGTCGAAGGCGGGACCGATAACTGG GGTGAAGTCGTAA

REFERENCES

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Although the presently disclosed subject matter and its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the present disclosure as defined by the appended claims. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps described in the specification. As one of ordinary skill in the art will readily appreciate from the disclosure of the presently disclosed subject matter, processes, machines, manufacture, compositions of matter, means, methods, or steps, presently existing or later to be developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein can be utilized according to the presently disclosed subject matter. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.

Patents, patent applications, publications, product descriptions and protocols are cited throughout this application the disclosures of which are incorporated herein by reference in their entireties for all purposes. 

1.-54. (canceled)
 55. A method for determining susceptibility of an intestinal disorder in a companion animal, comprising: a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal; b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and c) determining that the companion animal is susceptible of an intestinal disorder, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism.
 56. The method of claim 55, wherein the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HQ802983.1.1440, GQ449092.1.1375, GQ448744.1.1393, KF842598.1.1394, HG798451.1.1400, New.ReferenceOTU52, HK555938.1.1357, FJ957494.1.1454, FN667392.1.1495, New.ReferenceOTU54, HQ760911.1.1437, GQ006324.1.1342, FJ950694.1.1472, FM865905.1.1392, FJ506371.1.1371, FJ957528.1.1445, JF712675.1.1540, New.ReferenceOTU82, AB009242.1.1451, HQ751549.1.1448, AB506370.1.1516, DQ057365.1.1393, FN667422.1.1495, AJ270486.1.1241, FN668375.4306350.4307737, GQ867426.1.1494, GX182404.8.1529, JF224013.1.1362, GQ448246.1.1389, KC245406.1.1465, FN667084.1.1493, EU470512.1.1400, EU768569.1.1352, AY239462.1.1500, KC504009.1.1465, FM179752.1.1686, New.ReferenceOTU114, HK557089.3.1395, JQ208181.1.1352, HQ803964.1.1435, AM276759.1.1484, JN387556.1.1324, GQ448486.1.1387, HK694029.9.1487, HQ754680.1.1441, FN563300.1.1447, FP929060.3837.5503, GQ448506.1.1374, Enterococcus durans, C. perfringens, or E. coli.
 57. The method of claim 56, wherein the first intestinal microorganism is C. perfringens, E. coli and any combination thereof.
 58. The method of claim 55, wherein the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of EU774020.1.1361, HQ793763.1.1451, HQ792787.1.1438, New.ReferenceOTU109, HQ792778.1.1436, or DQ113765.1.1450.
 59. The method of claim 55, further comprising providing a customized recommendation of a treatment regimen, and/or further monitoring the intestinal microorganism, when the first amount of the first intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.
 60. A method for determining responsiveness of a companion animal having an intestinal disorder to a diet, comprising: a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal; b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and c) determining that the companion animal is responsive to the diet, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism, or determining that the companion animal is non-responsive to the diet, when the first amount of the intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.
 61. The method of claim 60, wherein the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of New.ReferenceOTU45, JRPJ01000002.1034290.1035971, KF842598.1.1394, JF920309.1.1340, FJ978526.1.1378, New.ReferenceOTU54, HQ793763.1.1451, DQ113765.1.1450, DQ797046.1.1403, ACBW01000012.3536.5054, JN387556.1.1324, New.ReferenceOTU52, or JQ208053.1.1336.
 62. The method of claim 60, wherein the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK693629.1.1491, GQ493166.1.1359, GQ491426.1.1332, FJ957494.1.1454, GQ449092.1.1375, GQ448486.1.1387, AMCI01001631.34.1456, or HK555938.1.1357.
 63. The method of claim 60, further comprising administering the diet to the companion animal when companion animal is determined as responsive to the diet.
 64. The method of claim 60, further comprising administering the diet, a steroid and optionally an antibiotic to the companion animal when companion animal is determined as non-responsive to the diet.
 65. The method of claim 60, wherein the determination in step c) occurs before administering the diet or the diet, the steroid and optionally the antibiotic to the companion animal.
 66. A method for determining effectiveness of a diet for treating an intestinal disorder in a companion animal, comprising: a) measuring a first amount of a first intestinal microorganism and/or a second amount of a second intestinal microorganism in the companion animal before or after administering a diet to a companion animal for treating an intestinal disorder; b) comparing the first amount of the first intestinal microorganism with a first reference amount of the first intestinal microorganism, and/or comparing the second amount of the second intestinal microorganism with a second reference amount of the second intestinal microorganism, wherein the reference amounts of the intestinal microorganisms are determined based on the amounts of the intestinal microorganisms in a plurality of healthy companion animals; and c) determining that the diet is effective for treating an intestinal disorder, when the first amount of the intestinal microorganism is higher than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is lower than the second reference amount of the second intestinal microorganism, or determining that the diet is ineffective for treating an intestinal disorder, when the first amount of the intestinal microorganism is lower than the first reference amount of the first intestinal microorganism, and/or when the second amount of the second intestinal microorganism is higher than the second reference amount of the second intestinal microorganism.
 67. The method of claim 66, wherein the first intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of HK557089.3.1395, or GQ448336.1.1418.
 68. The method of claim 66 or 67, wherein the second intestinal microorganism comprises one or more bacterium comprising a 16S rRNA comprising a nucleotide sequence that is at least about 90% homologous or identical to the 16S rRNA nucleotide sequence of KF842598.1.1394, GQ006324.1.1342, HQ802983.1.1440, JN387556.1.1324, FJ950694.1.1472, HG798451.1.1400, New.ReferenceOTU52, or GQ448468.1.1366/
 69. The method of claim 66, further comprising administering the diet to the companion animal when companion animal is determined as responsive to the diet.
 70. The method of claim 66, further comprising administering the diet, a steroid and optionally an antibiotic to the companion animal when companion animal is determined as non-responsive to the diet.
 71. The method of claim 66, wherein the determination in step c) occurs before administering the diet or the diet, the steroid and optionally the antibiotic to the companion animal. 72.-94. (canceled)
 95. The method of claim 60, wherein the diet comprises a food product comprising an effective amount of a bacterium capable of producing a first bile acid, wherein the bacterium is C. hiranonis.
 96. The method of claim 66, wherein the diet comprises a food product comprising an effective amount of a bacterium capable of producing a first bile acid, wherein the bacterium is C. hiranonis. 